production equipment for photovoltaics - singulus

Review ArticleCDKCCN and CDKI Alterations forCancer Prognosis and Therapeutic Predictivity

Patrizia Bonelli Franca Maria Tuccillo Antonella BorrelliAntonietta Schiattarella and Franco Maria Buonaguro

Molecular Biology and Viral Oncology Unit Department of Research Istituto Nazionale Tumori-IRCCS Fondazione ldquoG Pascalerdquo80131 Naples Italy

Correspondence should be addressed to Patrizia Bonelli pbonelliistitutotumorinait

Received 9 October 2013 Accepted 4 December 2013 Published 29 January 2014

Academic Editor Maria Lina Tornesello

Copyright copy 2014 Patrizia Bonelli et alThis is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The regulation of cell growth and division occurs in an accurate sequential manner It is dictated by the accumulation of cyclins(CCNs) and cyclin-dependent kinases (CDKs) complexes and degradation of CCNs In human tumors instead the cell cycle isderegulated causing absence of differentiation and aberrant cell growth Oncogenic alterations of CCNs CDKs and CDKIs havebeen reported in more than 90 of human cancers and the most frequent are those related to the G1 phase Several molecularmechanisms including gene overexpression chromosomal translocations point mutations insertions and deletions missense andframe shift mutation splicing or methylation may be responsible for these alterations The cell cycle regulators are involved intumor progression given their association with cancers characterized by higher incidence of relapses and chemotherapy resistanceIn the last decade anticancer drug researches focused on new compounds able to target molecules related to changes in genesassociated with tumor status Recently the studies have focused on the restoration of cell cycle control modulating moleculartargets involved in cancer-cell alterations This paper aims to correlate alterations of cell cycle regulators with human cancers andtherapeutic responsivity

1 Introduction

The recent progress in the field of molecular medicine hasidentified several molecular markers involved in the regula-tion of the cell cycle as a target for prognosis and cancer treat-ment Cell cycle is deregulated in human tumors causing theabsence of differentiation and aberrant cell growth [1ndash3]Thecell cycle includes cell division differentiation growth andprogrammed cell death through apoptosis The regulationof this process involves environmental stimuli that lead tothe activation of cyclin-dependent serinethreonine kinases(CDKs) regulated by cyclins (CCNs) and inhibitors of cyclin-dependent kinases (CDKIs) The main phases regulated byCDKs are the DNA integrity control checkpoints mediatedby the retinoblastoma susceptibility gene suppressor (Rb) thetumor suppressor gene TP53 and transcription factors of theE2F family [4 5]

So far nine CDKs and at least 15 CCNs have been identi-fied [6ndash8] The CDKs proteins of 300 amino acids in length

are activated by a no covalent binding with specific cyclinstriggering the transition between different phases of the cellcycle The formation of the complex CCNCDK is usuallytransient and is regulated by ubiquitin-mediated degradationThe CDKs are negatively regulated by endogenous inhibitorsCDKIs So far two families of inhibitors have been identifiedthe p21 and the p16 families The p21 family includes p21CDKN1A p27CDKN1B and p57CDKN1C [9 10] The p16family includes p16CDKN2A p15CDKN2B p18CDKN2Cand p19CDKN2D [9]The p21 family members interact withboth CCNs and CDKs subunit while members of the p16family interact only with the CDKs [7]

2 Cell Cycle Regulators

In Figure 1 an overview of the mammalian cell cycle isreported The regulation of cell growth and division occurs

Hindawi Publishing CorporationBioMed Research InternationalVolume 2014 Article ID 361020 15 pageshttpdxdoiorg1011552014361020

2 BioMed Research International

in a precise sequential manner It is dictated by the accumu-lation of CDKCCN complexes and degradation of CCNsQuiescent cells in G0 enter the G1 phase in response toexternal stimuli such as mitogenic growth factors or on thebasis of internal needs Cyclin D (CCND) binds to andactivates CDK4 andor CDK6 depending on the cell typeThe complexCCNDCDK4 orCDK6 phosphorylates RbThecyclin E (CCNE) binds to and activates CDK2 resulting inphosphorylation of a different site on Rb The E2F transcrip-tion factors (EF1ndashEF5) dissociate from the hyperphosphoryl-ated form of Rb to activate the transcription of genes thatpromote S phase including the thymidylate synthase (TS) thedihydrofolate reductase (DHFR) and the DNA polymerase(POL) [11 12] From this point on the cell has passed theldquorestriction pointrdquo and becomes committed to the progres-sion in the cell cycle and independent of growth factorsCCNA and CCNE bind to CDK2 allowing the cell to pro-ceed through the S phase The complex CCNACDK2 facil-itates the transition from the S phase to the G2 phase Thecomplexes CCNBCDK1 accumulate in late G2 phase neces-sary for the progression of the cell through the M phase [13]Following completion of anaphase CCNB is degraded thusreturning the cell to G1 state which in the presence of stim-ulation by growth factors proceeds by successive cycles ofcell division CDC25 (A B and C) are also required for theactivation of CDK complexes that control progressionthrough the cell cycle Activation of CDKs can be achievedthrough dephosphorylation bymembers of the CDC25 phos-phatase family (CDC25A CDC25B andCDC25C) CDC25Aplays an important role at theG1S-phase transition CDC25Bundergoes activation during S phase and plays a role inactivating the mitotic kinase CDK1CCNB in the cytoplasmActive CDK1CCNB then phosphorylates and activatesCDC25C leading to a positive feedback mechanism and toentry into mitosis [14] The repair of DNA damage or apop-tosis occurs prevalently at checkpoint G1 while the integrityof the synthesizedDNA is examined atG2 to ensure the fidel-ity of the replicated genome [15 16]TheCDKs play an impor-tant role in the regulation of these checkpoints For examplein response to various stress signals TP53 a transcriptionfactor is activated and causes the transcriptional inductionof CDKN1A and the cell cycle arrest at the G1 checkpoint[10] The length of the individual phases of the cell cycle canvary depending on the cell type and the particular conditionsThe activity of CDKs during the cell cycle is controlled atmultiple levels including the association with CCN acti-vating transient expression and rapid degradation of CCNsposttranslational modifications by kinases and phosphatasesinteractions with CDKIs and intracellular translocations [9]

3 Cell Cycle in Cancer

Oncogenic alterations of CCNs CDKs CDKIs and othercomponents of Rb pathway have been reported in more than90 of human cancers [1 2 17ndash40] as summarized in Table 1

One of the most relevant alterations is represented bythe p16 (CDKN2A)-CCNDsCDK-Rb pathway frequentlyaltered in various types of cancers [41ndash43] In solid tumors

DNA synthesis (S)

Cyclin BCDK1

Cyclin ACDK1

Cyclin DCDK4 CDK6

Cyclin ECDK2

Cyclin ACDK2

G2 G1

Mitosis (M)

Mammaliancell cycle

Figure 1 The mammalian cell cycle Cyclins and cyclin-dependentkinases are referred to their specific phase of the cell cycle Followingmitogenic signals that promote the cell entry in the G1 phasethe progression through the cell cycle is regulated by sequentialactivation of cyclins and cyclin-dependent kinases

there is a correlation between genetic alterations within thispathway and clinical outcome in cancer patients [44] Anom-alies of CDKs and CCNDs are very frequent in the G1 phaseSeveral molecular mechanisms may be responsible for thesealterations including gene overexpression chromosomaltranslocations pointmutations insertions and deletionsmis-sense and frame shift mutation splicing or methylationCDKs are found overexpressed in several types of tumorsincluding sarcoma colon carcinoma and lymphoma [45ndash47] CDK overexpression can be caused by gene amplification[48] chromosome translocation or point mutations [49]that impair the kinase interaction with CDK inhibitors asthe case for CDK4 in some melanoma patients [50] CDKactivity can also be dysregulated by overexpression of thecyclin partner or inactivation of CDK inhibitors both eventsbeing quite common in tumors [51 52]The lack of regulationof CDKs by its inappropriate activation is essential for main-taining the malignant transformation Changes of CCND1gene expression have been reported in several neoplasias Inparticular CCND1 gene is induced (transactivation) by var-ious oncogenic signals including the activating mutationof ras genes src and mitogen-activated protein kinases(MAPK) [53 54] as well as myc [55 56] Moreover chromo-somal aberrations involving CCND1 have been reported inB-lymphocytic malignancy and multiple myeloma [57 58]CCND1 overexpression played a role in the pathogenesis ofmammary cancer in transgenic mice [59 60] and lymphoma[61]The dysregulation of CCNE is associated with hyperpro-liferation and malignant transformation [26] Overexpres-sion of CCNE1 has been linked to endometrial hyperplasiaandor carcinoma [25] CCNE1 is overexpressed in manyhuman tumors in particular breast cancer and also nonsmall

BioMed Research International 3

Table 1 Alterations of regulators of cell cycle cyclins cyclin-dependent kinases and cyclins-dependent kinases inhibitors prevalent inhuman cancers

Regulators of cell cycle Cycle phase or activity Tumors

CCND1 G1Lymphoma (90) breast (50) and lung carcinoma (15)sarcoma (30) hepatocellular carcinoma (20) urothelialcarcinoma (14) cervical carcinoma (24)

CCND2 G1 CLL colorectal carcinoma lung carcinoma (20)

CCND3 Late G1 early S Lymphoma (50) retinoblastoma urothelial carcinoma (11)

CCNE G1SGastric and colorectal carcinoma breast carcinoma pancreaticcarcinoma bladder carcinoma (50)

CCNB1 G2M Colorectal carcinoma breast carcinoma thyroid carcinoma (19)

CCNA SG2 Breast carcinoma hepatocellular carcinoma

CDK2 G1S Colorectal carcinoma

CDK4 G1SMelanoma colorectal carcinoma breast carcinoma oral squamouscarcinoma (50) cervical carcinoma (26)

CDK6 G1S Glioma melanoma oral squamous carcinoma (40)

CDKN2B Inhibition of CDK4CDK6ALL (35) lung carcinoma (35) melanoma urothelialcarcinoma (23)

CDKN2A Inhibition of CDK4CDK6Melanoma glioma breast carcinoma nasopharyngeal carcinomaurothelial carcinoma (23)

CDKN1A Inhibition of all CDKs Melanoma leukemia colorectal carcinomaCDKN1B Inhibition of all CDKs Melanoma breast carcinoma colon carcinoma

cell lung cancer leukemia and others [62] CCNE has beenfound to be amplified overexpressed or both in some casesof breast and colon cancer and in acute lymphoblastic andmyeloid leukaemia [63ndash65]

4 Clinical Implication of CellCycle Dysregulation

41 Cell Cycle and Cancer Prognosis The cell cycle regulatorsas CCNs and CDKIs are involved in the mechanisms oftumor progression CCND is associated with higher inci-dence of relapses in tumors of the head and neck [66] andin chemotherapy resistance [67] Tumors that overexpressCCND1 generally have a poor prognosis [68ndash70] Also over-expression of CCNE has been reported to be a poor prog-nostic factor in cancers of various organs [71ndash73] Transgenicmice overexpressing human CCNE spontaneously developedmammary carcinoma [74] CCNE overexpression correlateswell with the aggressiveness of breast cancer [75] with gastriccancer progression [76] and is predictive of the risk of dis-tant recurrence in the abdomen [77] The inactivation ofendogenous inhibitors of p16 or p21 family due to theirmuta-tiondeletion or TP53-mediated changes causes aberrantactivity of CDK and inactivation of RbThe loss of CDKN2ACDKN1B and CDKN1A is a predictor of poor prognosisin several types of cancer [78ndash83] The loss of CDKN2Aappears to be closely related to the functional inactivation ofCDKN1B and assessment of CDKN1A status may be usefulfor a precise prognostic prediction of individuals with HCC

expressing high levels of CDKN1B [84] Hypermethylation ofCDKN1A promoter suppresses CDKN2A expression with asubsequent poor prognosis in patients with esophageal squa-mous cell carcinomas [85] Loss of CDKN1B was associatedwith poor prognosis in patients with Dukesrsquo B tumor or thosewith proximal tumor [80] and in patients with pancreaticcancer [81] Tenjo et al [82] observed that altered CDKN1Bexpression was a predictor of poor prognosis for patientswith stage III colorectal cancers Codeletion of CDKN2BCDKN2A genes is significantly related to the prognosis ofNSCLC patients whereby detecting codeletion of both genesmight be used as a potential marker for NSCLC prognosis[83] The CDKN2ACDKN2B deletion correlates with a highrisk of relapse or death in patients with ALL [86 87] Myelo-dysplastic syndromes patients with CDKN2B gene methyla-tion at diagnosis or in subsequent studies had a significantlyhigher chance of disease progression toAML than thosewith-out the gene methylation [88] The CDKN1B protein nega-tively regulates G1 progression by binding to G1 CCNCDKcomplexes and inhibits their activity resulting in inhibitionof entry to the cell cycle Reduced levels of CDKN1B occur inseveral cancer types and are generally associated with poorprognoses For example loss ofCDKN1B has been revealed tobe an independent prognostic factor in breast colon and gas-tric carcinomas [89 90] Gastric tumors with high CDKN1Bwerewell differentiated with low levels of invasion and lymphnode metastasis CDKN1B-negative cases demonstrated apoor prognosis [91] Expression of CDKN1B is significantlydecreased in renal cell carcinoma (RCC) as compared with


Table 2 Cell cycle modulators and their molecular target optimaltherapeutic inhibitors

Agent TargetStaurosporine CDK1 CDK2 CDK4Flavopiridol CDK1 CDK2 CDK4 CCNDButyrolactone CDK1 CDK2Paullones CDK1 CDK2 CDK5Indirubin CDK1 CDK2 CDK4 CDK5Rapamycin CCND CCNAOlomoucine CDK1 CDK2 CDK5Isopentenyladenine CDK1 CDK2

normal kidney tissue Loss of CDKN1B expression is a riskfactor for disease recurrence and the strongest predictor ofcancer-specific survival [92]

The expression of CDKN1A gene acts as an inhibitor ofthe cell cycle during G1 phase and is tightly controlled by thetumor suppressor protein TP53 Normal cells generally dis-play a rather intense nuclear CDKN1A expression Loss ofCDKN1A expression has been associated with poor prognosisin several carcinomas [93]

Recently it has been demonstrated that microRNAs(miRNAs) a class of small noncoding RNAs control the reg-ulators of cell cycle modulating their gene expression miR-24 directly targets CDKN1B and CDKN2A in keratinocytesand in different cancer-cell lines promoting their prolifera-tion It is involved in posttranscriptional regulation of CDKIsand its upregulation may play a role in carcinogenesis [94]Several studies showed that CDKN1A could be regulated atthe translational level by miRNA such as miR-93 miR-20abmiR-17 andmiR-106ab [95 96]Wu et al have demonstratedthat CDNK1A can be directly targeted and modulated bymultiple miRNA molecules [97]

42 Cell Cycle andTherapeutics Although chemotherapeuticdrugs save many lives they are very toxic nonselectiveand less effective than desired In the last decade anticancerdrug researches focused on new compounds able to targetmolecules related to changes in genes associated with tumorstatus Recently the studies have focused on the restorationof cell cycle control modulating molecular targets involvedin cancer-cell alterations Several potential strategies havebeen proposed such as inhibition of CDKs downregulationof cyclins overexpression of endogenous CDKIs disruptionof the interactions CCNCDK altered proteolysis degra-dation of CCNs and specific inhibition of tyrosine kinaseleading to activation of the cell cycle In Table 2 severalcompounds have been reported according to their moleculartargets The response of the tumors to modulators of cellcycle varies from simple cytostasis to cell death depending onthe concentrations used and the downstream result of the cellcycle arrest 7-Hydroxystaurosporine (UCN-01) has shownantitumor activity against several human cancer-cell linesUCN-01 inhibits the cell cycle progression from the G1 to theS phase and is associated with inhibition of cyclin-dependent

kinase (CDK) activity and induction ofCDKN1AThe combi-nation of UCN-01 and tamoxifen results in augmented cyto-toxicity and may have a potential clinical application in thetreatment of breast cancer [98] Butyrolactone 1 inhibitor ofCDKI accelerates CCNB1 accumulation in G2M of renalcells which shifted to G1 phase without cell division [99]Indirubin-31015840-oxime (I3M) an indigo alkaloid was found todisplay potent antitumor activities on various types of cancercells I3M increased the level of CDKN1B and reduced thelevels of CDK2 and CCNE in neuroblastoma cells arrestingthe cells at G0G1 phase [100] Many indirubin derivativeshave been studied for their potential antisolid tumor activity[101] Despite years of research and attempts directed at inhib-iting cell cycle kinases or cell cycle regulating transcriptionfactors most of these approaches have not been successfullytranslated to the clinic as cancer therapeutics [102]

However the inhibition of CDKs is particularly attractivefrom the standpoint of cancer given their key role in the cellcycle High throughput screening and drug design based onthe structure have produced several new compounds thatinhibit the activity of CDKs in very specific manner Cyclin-dependent kinases (CDKs) control cell cycle progressionRNA transcription and apoptosis making them interestingtargets for anticancer drug development A promising pyra-zolo[1 5-a] pyrimidine compound devoid of ABC trans-porter interaction has been identified as a highly suitabledrug for further preclinical and clinical evaluation in cancertreatment [103] Tanshinone IIA (Tan-IIA) is one of themajorlipophilic components isolated from the root of Salviae Mil-tiorrhizae radix Chiu et al [104] have explored the mecha-nisms of cell death induced by Tan-IIA treatment in prostatecancer cells in vitro and in vivoTheG0G1 phase arrest corre-lated with increase of CDK inhibitors (CDKN2A CDKN1Aand CDKN1B) and decrease of the checkpoint proteins[104] The effects of euphol a tetracyclic triterpene alcoholisolated from the sap of Euphorbia tirucalliwere examined inT47D human breast cancer cells Treatment of the cellswith euphol resulted in decreased cell viability which wasaccompanied by an accumulation of cells in the G1 phaseEuphol treatment downregulated CCND1 expression and thehypophosphorylation of Rb Furthermore this effect was cor-elated with the downregulation of CDK2 expression and theupregulation of the CDKIs (CDKN1A and CDKN1B) as wellas reduced expression levels of CCNA and CCNB1 [105]Treatment options for hepatocellular carcinoma (HCC) usingchemotherapeutics at intermediate and advanced stages ofdisease are limited as frequently HCC escape from therapyand patients succumb to disease progression The effective-ness of the novel compounds BA-12 andBP-14 that antagonizeCDK1257 and CDK9 has been studied in HCC patientssince CDKs aberrant activation is frequently observed insuch patients Inhibition of those CDKs in human HCC celllines reduced the clonogenicity arresting the cells in SG2 andG2M boundaries and inducing their apoptosis In vivo inmouse xenograft this treatment also inhibited tumor devel-opment [106] On the contrary dysregulation of CDKN1Bdue to proteolysis frequently results in tumorigenesis Novelcompounds that inhibited CDKN1B degradation have been


identifiedThese compounds inhibit CDKN1B ubiquitinationin vitro as well as its degradation in cell cultures [107] Anovel series of 2-substituted-6-biarylmethylamino-9-cyclo-pentylpurine derivatives has been synthesized and screenedfor improved CDK inhibitory activity and antiproliferativeeffects One of the most potent compounds 6b exhibitedstrong cytotoxicity in the human melanoma cell line G361that correlated with robust CDK1 and CDK2 inhibition andcaspase activation [108] Decorin a component of extracel-lular matrix has been involved in the inhibition of cell pro-liferation upregulating CDKN1A and reactivating CDKN1Cexpression in HepG2 tumor cells [109] CDK inhibitionmay be particularly successful in hematologic malignancieswhich are more sensitive to cell cycling inhibition and apop-tosis induction In general the antitumor efficacy of CDKinhibitor as monotherapy is modest and rational combi-nations are being explored including those involving othertargeted agents While selective CDK46 inhibition might beeffective against certain malignancies broad-spectrum CDKinhibition will likely be required for most cancers [110] Aspecific inhibitor of CDK46 has been developed recentlyPD-0332991 proved to be effective in the treatment of breastcancer [111] dFMGEN a novel genistein derivative is a can-didate for cancer therapy arresting the cell cycle at G1 phasewith significant reduction of CDK4 and CCND1 proteinlevels This reduction is caused by CDKN2B CDKN1A andCDKN1B level increase and the subsequent decrease of pro-tein levels directly suppressed Rb phosphorylation and E2F1activity [112] Studies of drug combination showed thatflavopiridol potentiated the cytotoxicity induced by theRaf inhibitor sorafenib This potentiation correlated withenhanced apoptosis and suppression of Rb signaling [113]Recently the research of new targets for cancer therapyfocused on long noncoding RNAs (lncRNAs) that are involvedin the regulation of critical regulators of cell cycle as CDKsCCNs and CDKIs [114] Human tumors generally exhibitaltered expression of miRNAs with oncogenic or tumor-suppressive activity miRNA-based cancer gene therapy offersthe theoretical possibility of targeting gene networks that arecontrolled by a single aberrantly expressed miRNA Inexperimental models the restoration of tumor-suppressivemiRNA or sequence-specific knockdown of oncogenic miR-NAs by ldquoantagomirsrdquo has produced promising antitumor out-comes [115 116]

All such data clearly show the relevance of such anapproach for cancer treatment and the need to identify theappropriate combination with conventional anticancer ther-apy

5 Nonsteroidal Anti-Inflammatory Drugs asModulators of the Cell Cycle in Cancers

Nonsteroidal anti-inflammatory drugs (NSAIDs) are prima-rily used as analgesics for the relief of pain and to controlinflammation [117] NSAIDs inhibit cyclooxygenase (COX)activity and the synthesis of prostaglandins which are medi-ators of inflammation Numerous epidemiological clinical

and laboratory studies have also suggested that NSAIDsinhibit the promotion and proliferation of some tumors [118ndash120] However the antiproliferative activity of NSAIDsrequires concentrations that are 100- to 1000-fold higherthan the concentrations necessary to inhibit COX activity[121] Induction of COX-2-independent apoptosis has beendescribed in HT29 colon cancer cells Apoptosis was inducedin these cells by the inhibition of 3-phosphoinositidedepend-ent kinase 1 (PDK1) [122]Moreover apoptosis and a cell cycleblockadewere observed inHCT-15 colon carcinoma cells thatexpressed only COX-1 (and not COX-2) [123] NO-aspirin-induced cell cycle arrest and apoptosis of pancreatic cancercells have been shown to occur via ROS-mediated mod-ulation of all three MAP kinase signaling pathways andtheir downstream effector molecules such as CDKN1A andCCND1 [124] Celecoxib has been shown to inhibit cancergrowth independently of COX-2 expression levels with a G0G1 cell cycle arrest and decreased levels of CCNA andCCNB1or CCND1 depending on tumor type [125 126] NSAIDsincreased CDKN1B by inhibiting protein degradation to sup-press the proliferation of human lung cancer cells [127]Salicylates inhibit the proliferation through upregulation ofthe CDKN1B and CDKN1A This was associated with adecrease inCDK2 and to a lesser extent inCDK6 activity thuspreventing hyperphosphorylation of Rb and cell cycle pro-gression Similar to salicylates however sulindac reduced theproliferation rate of HT-29 colon carcinoma cells and causedthem to accumulate in the G0G1 phase This was associatedwith reduced expression and reduced catalytic activity ofcyclin-dependent kinases (CDK1 CDK2 and CDK4) [128]These data strongly suggest the need to use such moleculesas complementary therapeutic drugs besides their relevantchemopreventive role

51 Changes of the Cell Cycle Gene Expression Profile of GastricCancer Cells in Response to Ibuprofen An Anticancer ModelThe primary purpose of the studies performed in our labo-ratory was to verify the antitumor effects of ibuprofen a com-monly used NSAID on the MKN-45 human gastric cancer-cell line [129] where an inhibitory effect of the ibuprofen oncell proliferation was observed These results were in agree-ment with data obtained in vitro using other tumor cell lines[130 131] as well as in vivo on xenografts of MKN-45 cells[132] Ibuprofen was used at concentrations ranging between400 and 800 120583M and the cell proliferation rate was signifi-cantly reduced in a time- and concentration-dependentman-ner Previous studies showed that concentrations of ibuprofencomparable to those used in our studies targeted a widevariety of cellular processes [122 133ndash135] Moreover theysuggested the existence of other targets or COX-independentmechanisms that could be responsible for the antiprolifera-tive effects of ibuprofen Our data have shown that ibuprofenis involved in the cell cycle arresting the cells in active rep-lication at the G0G1 phase Ibuprofen treatment in factcaused MKN-45 cells to shift from the S and G2M phasesto the G0G1 phase resulting in a significant accumulation ofcells at the resting phase as previously reported on differentcell lines [133 136ndash138] The cell cycle changes caused in


Table3Fu

nctio

nalgroup

ingof

genesu

p-anddo

wnregulated

after

ibup

rofentre

atment

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

MCellcyclecheckp

oint

G1S

DNAdamage

checkp

oint

CDC2

5ANM

001789

mdashmdash

minus2090

mdashmdash

mdashminus240

8mdash

mdashCelld

ivision

cycle

25ho

molog

A(Spom

be)

CDC2

5BNM

021873

mdashmdash

mdashmdash

mdashmdash

minus2980minus2686minus2415

Celld

ivision

cycle

25ho

molog

B(Spom

be)

CDC2

5CNM

001790

mdashmdash

minus2155minus2306

mdashminus2050minus3588

mdashmdash

Celld

ivision

cycle

25ho

molog

C(Spom

be)

p53-depend

entG

1SDNADam

age

checkp

oint

TP53

NM

000546

mdash244

72503

mdashmdash

mdash3150

mdash3650

Tumor

proteinp53

CDKN

1ANM

078467

mdash3074

2789

mdashmdash

mdashmdash

minus2254minus2203

Cyclin-depend

entk

inase

inhibitor1A(p21C

ip1)

GADD45A

NM

001924

mdashmdash

mdashmdash

mdashmdash

4194

2493

2047

Growth

arrestand

DNA-

damage-indu

ciblealph

a

GADD45B

NM

015675

2352

mdash2191

2864

2792

2899

4016

5479

5200

Growth

arrestand

DNA-

damage-indu

ciblebeta

GADD45G

NM

006705

mdash2208

2661

2379

4796

5081

1412

12253

2Growth

arrestand

DNA-

damage-indu

ciblegamma

TP53IN

P1NM

033285

2033

3475

5494

3071

mdashmdash

mdashmdash

mdashTu

mor

protein53

indu

cible

nucle

arprotein1

TP53I3

NM

004881

mdashmdash

mdashmdash

mdash2214

mdashmdash

240

6Tu

mor

proteinp53indu

cible

protein3

MDM2

NM

002392

2534

mdash2868

3019

4111

3165

mdash4369

mdashMdm

2p53bind

ingprotein

homolog

(mou

se)


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

MCellcyclemito

ticG1S

transition

CCNE1

NM

001238

mdashmdash

mdashmdash


mdashmdash

CyclinE1

CDKN

1ANM

078467

mdash3074

2789

mdashmdash

mdashmdash

minus2254minus2203

Cyclin-depend

entk

inase

inhibitor1A(p21C

ip1)

CDKN

1BNM

0040

64mdash

mdashmdash

mdashmdash

2477

2432

3367

4938

Cyclin-depend

entk

inase

inhibitor1B(p27K

ip1)

CDC2

NM

001786

mdashmdash

minus2540minus3260minus2250minus3940minus5820minus2730minus3420

Celld

ivision

cellcycle

2RB

1NM

000321

mdashmdash

mdashmdash

mdashmdash

mdash2872

2473

Retin

oblasto

ma1

DNAreplicationE

2Fmediatedregu

latio

nof

DNAreplication

E2F1

NM

005225

mdashmdash

mdashminus2283minus3073minus2740minus75

52minus4090minus3215

E2Ftranscrip

tionfactor

1

CDT1

NM

030928

mdashmdash

mdashminus2571minus2727minus2332minus5931minus3269minus2202

Chromatin

licensin

gandDNA

replicationfactor

1PC

NA

NM

002592

mdashmdash


Proliferatin

gcellnu

clear

antig

enDHFR

NM

000791

mdashmdash

mdashmdash

minus246

6minus2157minus2376minus4963minus77

12Dihydrofolateredu

ctase

TYMS

NM

001071

mdashmdash


95minus4872minus8869minus99

76minus11282

Thym

idylates

ynthase

POLA

2NM

002689

mdashmdash

mdashmdash

mdashmdash


DNApo

lymerasea

lpha2

POLA

1NM

016937

mdashmdash

mdashmdash

mdashmdash

minus2098

mdashminus2014

DNApo

lymerasea

lpha

1


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

POLD

2NM

006230

mdashmdash

mdashmdash

minus2794minus2403minus2611minus3001minus2114

DNApo

lymerase

delta

2

POLD

3NM

006591

mdashmdash

minus2244

mdashminus2135

mdashminus2992

mdashmdash

DNApo

lymerase

delta

3PO

LE2

NM

002692

mdashmdash


DNApo

lymerasee

psilo

n2

POLE

4NM

019896

mdashmdash

mdashmdash

mdashminus2154

mdashminus2118

mdashDNApo

lymerasee

psilo

n4

ORC

1LNM

004153

mdashmdash

mdashminus2914minus2588minus2538minus5605minus2337

mdashOrig

inrecogn

ition

complex

subu

nit1-like

(yeast)

RRM2

NM

001034

mdashmdash

mdashminus246

7minus2485minus4266minus6902minus4767minus4299

Ribo

nucle

otider

eductase

M2

subu

nit

Regu

latio

nof

DNA

replication

ORC

2LNM

006190

mdashmdash

mdashmdash

mdashmdash

2288

2727

2570

Orig

inrecogn

ition

complex

subu

nit2-like

(yeast)

ORC

4LNM

002552

mdashmdash

mdashmdash

mdash3216

2171

mdashmdash

Orig

inrecogn

ition

complex

subu

nit4

-like

(yeast)

ORC

5LNM

181747

mdashmdash

mdashmdash

mdashmdash

2209

2902

2765

Orig

inrecogn

ition

complex

subu

nit5-like

(yeast)

ORC

6LNM

014321

mdashmdash

mdashminus2348

mdashmdash

mdashminus2415

mdashOrig

inrecogn

ition

complex

subu

nit6

-like

(yeast)

MCM

2NM

004526

mdashmdash

mdashminus266


Minichrom

osom

emaintenance

complex

compo

nent

2

MCM

3NM

002388

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

3

MCM

4NM

005914

mdashmdash

mdashminus2267minus2266minus246

4minus3923minus504

0minus73

44Minichrom

osom

emaintenance

complex

compo

nent

4

MCM

5NM

006739

mdashmdash



Minichrom

osom

emaintenance

complex

compo

nent

5


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

MCM

6NM

005915

mdashmdash


mdashmdash

mdashmdash

Minichrom

osom

emaintenance

complex

compo

nent

6

MCM

7NM

182776

mdashmdash

minus2175minus2563

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

7

TK1

NM

003258

mdashmdash


6minus75

57minus24478minus75

67minus5991

Thym

idinek

inase1solub

le

FEN1

NM

004111

mdashmdash

minus2008minus2268

mdashminus2110


Flap

structure-specific

endo

nucle

ase1

LIG1

NM

000234

mdashmdash


4minus6958minus564

0DNALigase

1

Apop

tosis

CASP

3NM

004346

mdashmdash

mdashmdash

mdash2097

mdash2239

2112

Caspase-3

CASP

7NM

033339

mdashmdash

mdashmdash

2124

2097

2050

2335

2393

Caspase-7

CASP

8NM

033356

mdashmdash

mdashmdash

mdashmdash

3686

2420

3056

Caspase-8

CASP

9NM

001229

mdashmdash

mdashmdash

mdashmdash

mdash2221

2257

Caspase-9

CASP

10NM

032977

mdashmdash

mdashmdash

mdashmdash

mdash3617

4514

Caspase-10

DFFA

NM

213566

mdashmdash

mdashmdash

mdashmdash

mdashmdash

2034

DNAfragmentatio

nfactor

45kD

aalph

apolypeptid

e

DFF

BNM

001004

285

mdashmdash

mdashmdash

mdashmdash

4015

2121

4583

DNAfragmentatio

nfactor40

kDbetap

olypeptid

e(caspase-activated

DNase)


MKN-45 cells by ibuprofen correlated with alterations in cellcycle regulatory genes The subsequent microarray analysisrevealed that ibuprofen treatment for 24 h affected genesbelonging to ldquocell cyclerdquo pathways (Table 3) At 24 h most ofthe altered genes were G1S transition genes such as CDC25CDKN1A and TP53 CDC25 one of the key regulators of cellcycle transition [139] with protooncogenic properties andcarefully regulated at multiple levels [140 141] is downregu-lated at all timepoints (since the 24 h testing) in presence of allibuprofen concentrations including the lower 400 120583M doseCDKN1A gene instead is upregulated by more than 2-fold at24 h followed by downregulation at 72 h Its upregulationtightly controlled by TP53 is necessary to block cells in theG1phase and allow the repair of damaged DNA before theirentry into the subsequent S phase For such role CDKN1Arepresents the key effector of p53-dependent G0G1 phasearrest in response to different stress stimuli [142] TP53 levelsshow a similar significant upregulation for the whole ibupro-fen treatment period and to all tested doses with a late upreg-ulation likely associated with activation of apoptotic-relatedgenes The upregulation of TP53 signaling pathways in thefirst 24 h following ibuprofen treatment has been also shownby a previous study [133] TP53 signaling was activated early(24 h) as a result of intracellular modifications likely asso-ciated with oxidative stress which would justify the ROSincrease we observed In parallel to TP53 upregulation andcell cycle G0G1 arrest several S-related genes are downreg-ulated particularly those expressing proteins and enzymesinvolved in DNA replication Regulation of S-phase-criticalgenes is therefore an important component of G1 progres-sion to SThis coordinated regulation of S-phase genes is prin-cipally controlled by the E2F family of transcription factors[143] Treatment with ibuprofen for 48 h at all concentrationsdownmodulated S-phase-critical genes including E2F1 aswell as the E2F1-regulated genes and treatment for 72 hshowed amarkedmodulation of apoptosis-related transcripts(Table 3) Moreover the TP53-dependent G0G1 arrest wasparalleled with an increase in the expression of the cell cycleinhibitory protein CDKN1A whose induction is reported tobe transcriptionally upregulated by a TP53-dependent mech-anism Also the GADD45 gene was induced by activation ofthe TP53-dependent pathway and likely contributed to elicitgrowth arrest for an effective DNA repair or alternatively toinduce apoptosisThe ineffective repair of cell damage in factprevalently triggered in our model an apoptotic programcharacterized by upregulation of caspase transcripts in thelatter stages of the ibuprofen treatment

In conclusion this study showed that the early alterationsof cell cycle regulatory genes and the later induction of apop-tosis are the major mechanisms that account for the antipro-liferative effects of ibuprofen on the MKN-45 human gastriccancer-cell line Treatment with ibuprofen altered the cellcycle phase distribution by inhibiting the G1S transitionTheG0G1 arrest was associated with a decrease in the expressionof cyclins and cyclin-dependent kinases and an increase ofTP53 andCDKN1A alongwith downregulation of transcriptsencoding enzymes involved in DNA precursor synthesis andthe DNA replication system The prolonged ibuprofen treat-ment and the ineffective repair of damaged cells resulted in

the late upregulation of caspase transcripts with the conse-quent activation of an apoptotic program

Despite the high doses of ibuprofen required to elicit theapoptotic effects reported in our studies the implicatedmolecular mechanisms suggest that NSAIDs such as ibupro-fen may be of benefit in the treatment of cancers particularlyas local treatment

6 Conclusions

Several molecules and their pathways involved in the regu-lation of the cell cycle are currently considered as promisingprognostic biomarkers and target for cancer treatment Thishas been made possible by recent advances in moleculargenomic and proteomic technologies Characterization of thecell cycle pathways present in specific cancers will contrib-ute to improving diagnosis and cancer staging prognosticevaluation of cancer patients and optimal combinationalapproaches for innovative personalized treatment strategies

Disclosure

The authors have no relevant affiliations or financial involve-ment with any organization or entity with a financial interestin or financial conflict with the subject matter or materialsdiscussed in the paperThis includes employment consultan-cies honoraria stock ownership or options expert testimonygrants or patents received or pending or royalties Nowritingassistance was utilized in the production of this paper

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] C J Sherr ldquoCancer cell cyclesrdquo Science vol 274 no 5293 pp1672ndash1674 1996

[2] L H Hartwell andM B Kastan ldquoCell cycle control and cancerrdquoScience vol 266 no 5192 pp 1821ndash1828 1994

[3] M Dictor M Ehinger F Mertens J Akervall and J Wenner-berg ldquoAbnormal cell cycle regulation in malignancyrdquo AmericanJournal of Clinical Pathology vol 112 supplement 1 pp S40ndashS521999

[4] S van den Heuvel and E Harlow ldquoDistinct roles for cyclin-dependent kinases in cell cycle controlrdquo Science vol 262 no5142 pp 2050ndash2054 1993

[5] R A Weinberg ldquoThe retinoblastoma protein and cell cyclecontrolrdquo Cell vol 81 no 3 pp 323ndash330 1995

[6] G Draetta ldquoCell cycle control in eukaryotes molecular mech-anisms of cdc2 activationrdquo Trends in Biochemical Sciences vol15 no 10 pp 378ndash383 1990

[7] C J Sherr and J M Roberts ldquoCDK inhibitors positive andnegative regulators of G1-phase progressionrdquo Genes and Devel-opment vol 13 no 12 pp 1501ndash1512 1999

[8] E A Sausville J Johnson M Alley D Zaharevitz and A MSenderowicz ldquoInhibition of CDKs as a therapeutic modalityrdquo


Annals of the New York Academy of Sciences vol 910 pp 207ndash222 2000

[9] D O Morgan ldquoPrinciples of CDK regulationrdquo Nature vol 374no 6518 pp 131ndash134 1995

[10] Y Xiong G J Hannon H Zhang D Casso R Kobayashi andD Beach ldquop21 is a universal inhibitor of cyclin kinasesrdquoNaturevol 366 no 6456 pp 701ndash704 1993

[11] N Dyson ldquoThe regulation of E2F by pRB-family proteinsrdquoGenes and Development vol 12 no 15 pp 2245ndash2262 1998

[12] MHatakeyama andRAWeinberg ldquoThe role of RB in cell cyclecontrolrdquo Progress in Cell Cycle Research vol 1 pp 9ndash19 1995

[13] C J Sherr ldquoG1 phase progression cycling on cuerdquo Cell vol 79no 4 pp 551ndash555 1994

[14] I Nilsson and I Hoffmann ldquoCell cycle regulation by the Cdc25phosphatase familyrdquo Progress in Cell Cycle Research vol 4 pp107ndash114 2000

[15] L H Hartwell and T A Weinert ldquoCheckpoints controls thatensure the order of cell cycle eventsrdquo Science vol 246 no 4930pp 629ndash634 1989

[16] J K Buolamwini ldquoCell cycle molecular targets in novel anti-cancer drug discoveryrdquo Current Pharmaceutical Design vol 6no 4 pp 379ndash392 2000

[17] THunter and J Pines ldquoCyclins and cancer II cyclinD andCDKinhibitors come of agerdquo Cell vol 79 no 4 pp 573ndash582 1994

[18] J E Karp and S Broder ldquoMolecular foundations of cancer newtargets for interventionrdquoNature Medicine vol 1 no 4 pp 309ndash320 1995

[19] G F Draetta ldquoMammalian G1 cyclinsrdquo Current Opinion in CellBiology vol 6 no 6 pp 842ndash846 1994

[20] M H Dreyling L Bullinger G Ott et al ldquoAlterations of thecyclin D1p16-pRB pathway in mantle cell lymphomardquo CancerResearch vol 57 no 20 pp 4608ndash4614 1997

[21] M Hall and G Peters ldquoGenetic alterations of cyclins cyclin-dependent kinases and Cdk inhibitors in human cancerrdquoAdvances in Cancer Research vol 68 pp 67ndash108 1996

[22] F S Leach S J Elledge C J Sherr et al ldquoAmplification of cyclingenes in colorectal carcinomasrdquo Cancer Research vol 53 no 9pp 1986ndash1989 1993

[23] T Motokura and A Arnold ldquoCyclin D and oncogenesisrdquo Cur-rent Opinion in Genetics and Development vol 3 no 1 pp 5ndash101993

[24] M A Davis M Sturzl C Blasig et al ldquoExpression of humanherpesvirus 8-encoded cyclin D in Kaposirsquos sarcoma spindlecellsrdquo Journal of the National Cancer Institute vol 89 no 24pp 1868ndash1874 1997

[25] A L Niklaus M Aubuchon G Zapantis et al ldquoAssessmentof the proliferative status of epithelial cell types in the endo-metrium of young and menopausal transition womenrdquoHumanReproduction vol 22 no 6 pp 1778ndash1788 2007

[26] K Keyomarsi and T W Herliczek ldquoThe role of cyclin E in cellproliferation development and cancerrdquo Progress in Cell CycleResearch vol 3 pp 171ndash191 1997

[27] J M Gudas M Payton S Thukral et al ldquoCyclin E2 a novel G1cyclin that binds Cdk2 and is aberrantly expressed in humancancersrdquo Molecular and Cellular Biology vol 19 no 1 pp 612ndash622 1999

[28] M F Buckley K J E Sweeney J A Hamilton et al ldquoExpressionand amplification of cyclin genes in human breast cancerrdquoOncogene vol 8 no 8 pp 2127ndash2133 1993

[29] J Wang F Zindy X Chenivesse E Lamas B Henglein and CBrechot ldquoModification of cyclin A expression by hepatitis Bvirus DNA integration in a hepatocellular carcinomardquo Onco-gene vol 7 no 8 pp 1653ndash1656 1992

[30] J H Kim M J Kang C U Park et al ldquoAmplified CDK2 andcdc2 activities in primary colorectal carcinomardquoCancer vol 85no 3 pp 546ndash553 1999

[31] K Ikeda T Monden M Tsujie et al ldquoCyclin D CDK4 and p16expression in colorectal cancerrdquo Nippon Rinsho vol 54 no 4pp 1054ndash1059 1996

[32] J F Costello C Plass W Arap et al ldquoCyclin-dependent kinase6 (CDK6) amplification in human gliomas identified using two-dimensional separation of genomicDNArdquoCancer Research vol57 no 7 pp 1250ndash1254 1997

[33] A Kamb N A Gruis J Weaver-Feldhaus et al ldquoA cell cycleregulator potentially involved in genesis of many tumor typesrdquoScience vol 264 no 5157 pp 436ndash440 1994

[34] N Hayashi Y Sugimoto E Tsuchiya M Ogawa and Y Naka-mura ldquoSomatic mutations of the MTS (Multiple Tumor Sup-pressor) 1CDK4I (Cyclin-Dependent Kinase-4 Inhibitor) genein humanprimary non-small cell lung carcinomasrdquoBiochemicaland Biophysical Research Communications vol 202 no 3 pp1426ndash1430 1994

[35] P Cairns L Mao A Merlo et al ldquoRates of p16 (MTS1) muta-tions in primary tumors with 9p lossrdquo Science vol 265 no 5170pp 415ndash417 1994

[36] H G Drexler ldquoReview of alterations of the cyclin-dependentkinase inhibitor INK4 family genes p15 p16 p18 and p19 inhuman leukemia-lymphoma cellsrdquo Leukemia vol 12 no 6 pp845ndash859 1998

[37] S J Meltzer ldquoThe molecular biology of esophageal carcinomardquoRecent Results in Cancer Research vol 142 pp 1ndash8 1996

[38] H Takamura S Fushida T Hashimoto M Yagi and IMiyazaki ldquoAnalysis of the p16INK4 p15INK4B genes abnor-mality and the amplification of cyclin D1 gene in esophagealcancerrdquo Nippon Rinsho vol 54 no 4 pp 1043ndash1048 1996

[39] C Cordon-Cardo ldquoMutation of cell cycle regulators biologicaland clinical implications for human neoplasiardquo American Jour-nal of Pathology vol 147 no 3 pp 545ndash560 1995

[40] M Loda B Cukor S W Tam et al ldquoIncreased proteasome-dependent degradation of the cyclin-dependent kinase inhib-itor p27 in aggressive colorectal carcinomasrdquo Nature Medicinevol 3 no 2 pp 231ndash234 1997

[41] F Barbieri M Cagnoli N Ragni F Pedulla G Foglia and AAlama ldquoExpression of cyclin D1 correlates with malignancy inhuman ovarian tumoursrdquo British Journal of Cancer vol 75 no9 pp 1263ndash1268 1997

[42] T Zhang L B Nanney C Luongo et al ldquoConcurrent overex-pression of cyclin D1 and cyclin-dependent kinase 4 (Cdk4) inintestinal adenomas from multiple intestinal neoplasia (min)mice and human familial adenomatous polyposis patientsrdquoCancer Research vol 57 no 1 pp 169ndash175 1997

[43] KNakagawaNKConrad J PWilliams B E Johnson andMJ Kelley ldquoMechanism of inactivation of CDKN2 and MTS2 innon-small cell lung cancer and association with advancedstagerdquo Oncogene vol 11 no 9 pp 1843ndash1851 1995

[44] J Bartkova J Lukas P Guldberg et al ldquoThe p16-cyclin DCdk4-pRb pathway as a functional unit frequently altered inmelanoma pathogenesisrdquo Cancer Research vol 56 no 23 pp5475ndash5483 1996


[45] K Honoki H Fujii Y Tohma et al ldquoComparison of geneexpression profiling in sarcomas and mesenchymal stem cellsidentifies tumorigenic pathways in chemically induced rat sar-coma modelrdquo Oncology vol 2012 Article ID 909453 8 pages2012

[46] HYamamoto TMondenHMiyoshi et al ldquoCdk2cdc2 expres-sion in colon carcinogenesis and effects of cdk2cdc2 inhibitorin colon cancer cellsrdquo International Journal of Oncology vol 13no 2 pp 233ndash239 1998

[47] R Chiarle C Voena C Ambrogio R Piva and G InghiramildquoThe anaplastic lymphoma kinase in the pathogenesis of can-cerrdquo Nature Reviews Cancer vol 8 no 1 pp 11ndash23 2008

[48] M Marone G Scambia C Giannitelli et al ldquoAnalysis of cyclinE and CDK2 in ovarian cancer gene amplification and RNAoverexpressionrdquo International Journal of Cancer vol 75 no 1pp 34ndash39 1998

[49] J Easton T Wei J M Lahti and V J Kidd ldquoDisruption of thecyclin Dcyclin-dependent kinaseINK4retinoblastoma pro-tein regulatory pathway in human neuroblastomardquo CancerResearch vol 58 no 12 pp 2624ndash2632 1998

[50] L Zuo J Weger Q Yang et al ldquoGermline mutations in thep16(INK4a) binding domain of CDK4 in familial melanomardquoNature Genetics vol 12 no 1 pp 97ndash99 1996

[51] A J Obaya and J M Sedivy ldquoRegulation of cyclin-Cdk activityin mammalian cellsrdquo Cellular and Molecular Life Sciences vol59 no 1 pp 126ndash142 2002

[52] Y Matsuda ldquoMolecular mechanism underlying the functionalloss of cyclindependent kinase inhibitors p16 and p27 in hepat-ocellular carcinomardquoWorld Journal of Gastroenterology vol 14no 11 pp 1734ndash1740 2008

[53] R J Lee C Albanese R J Stenger et al ldquopp60(v-src) inductionof cyclin D1 requires collaborative interactions between theextracellular signal-regulated kinase p38 and Jun kinase path-ways a role for cAMP response element-binding protein andactivating transcription factor-2 in pp60(v-src) signaling inbreast cancer cellsrdquoThe Journal of Biological Chemistry vol 274no 11 pp 7341ndash7350 1999

[54] C Albanese J Johnson G Watanabe et al ldquoTransformingp21(ras) mutants and c-Ets-2 activate the cyclin D1 promoterthrough distinguishable regionsrdquo The Journal of BiologicalChemistry vol 270 no 40 pp 23589ndash23597 1995

[55] H Lovec A Grzeschiczek M Kowalski and T Moroy ldquoCyclinD1bcl-1 cooperates with myc genes in the generation of B-celllymphoma in transgenic micerdquo The EMBO Journal vol 13 no15 pp 3487ndash3495 1994

[56] J I Daksis R Y Lu L M Facchini W W Marhin and L J ZPenn ldquoMyc induces cyclin D1 expression in the absence of denovo protein synthesis and links mitogen-stimulated signaltransduction to the cell cyclerdquoOncogene vol 9 no 12 pp 3635ndash3645 1994

[57] C J de Boer J H J M van Krieken E Schuuring and P MKluin ldquoBcl-1cyclin D1 in malignant lymphomardquo Annals ofOncology vol 8 supplement 2 pp S109ndashS117 1997

[58] R Fiancette R Amin V Truffinet et al ldquoA myeloma translo-cation-like model associating CCND1 with the immunoglobu-lin heavy-chain locus 31015840 enhancers does not promote by itself B-cell malignanciesrdquo Leukemia Research vol 34 no 8 pp 1043ndash1051 2010

[59] T C Wang R D Cardiff L Zukerberg E Lees A Arnoldand E V Schmidt ldquoMammary hyperplasia and carcinoma inMMTV-cyclin D1 transgenic micerdquo Nature vol 369 no 6482pp 669ndash671 1994

[60] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005

[61] S E Bodrug B J Warner M L Bath G J Lindeman AW Harris and J M Adams ldquoCyclin D1 transgene impedeslymphocyte maturation and collaborates in lymphomagenesiswith the myc generdquoThe EMBO Journal vol 13 no 9 pp 2124ndash2130 1994

[62] T Moroy and C Geisen ldquoCyclin Erdquo International Journal ofBiochemistry andCell Biology vol 36 no 8 pp 1424ndash1439 2004

[63] K Keyomarsi D Conte W Toyofuku and M P Fox ldquoDeregu-lation of cyclin E in breast cancerrdquo Oncogene vol 11 no 5 pp941ndash953 1995

[64] P Schraml C Bucher H Bissig et al ldquoCyclin E overexpressionand amplification in human tumoursrdquoThe Journal of Pathologyvol 200 no 3 pp 375ndash382 2003

[65] H Lida M Towatari M Tanimoto Y Morishita Y Koderaand H Saito ldquoOverexpression of cyclin E in acute myelogenousleukemiardquo Blood vol 90 no 9 pp 3707ndash3721 1997

[66] R Michalides N van Veelen A Hart B Loftus E Wientjensand A Balm ldquoOverexpression of cyclin D1 correlates withrecurrence in a group of forty-seven operable squamous cellcarcinomas of the head and neckrdquo Cancer Research vol 55 no5 pp 975ndash978 1995

[67] E E Noel M Yeste-Velasco X Mao et al ldquoThe associationof CCND1 overexpression and cisplatin resistance in testiculargerm cell tumors and other cancersrdquo American Journal ofPathology vol 176 no 6 pp 2607ndash2615 2010

[68] S Gansauge F Gansauge M Ramadani et al ldquoOverexpressionof cyclin D1 in human pancreatic carcinoma is associated withpoor prognosisrdquo Cancer Research vol 57 no 9 pp 1634ndash16371997

[69] H Mineta K Miura S Takebayashi et al ldquoCyclin D1 overex-pression correlates with poor prognosis in patients with tonguesquamous cell carcinomardquoOralOncology vol 36 no 2 pp 194ndash198 2000

[70] T Sutter S Doi K A Carnevale N Arber and I B WeinsteinldquoExpression of cyclins D1 and E in human colon adenocarcino-masrdquo Journal of Medicine vol 28 no 5-6 pp 285ndash309 1997

[71] A Tzankov A Gschwendtner F Augustin et al ldquoDiffuse largeB-cell lymphoma with overexpression of cyclin E substantiatespoor standard treatment response and inferior outcomerdquo Clin-ical Cancer Research vol 12 no 7 I pp 2125ndash2132 2006

[72] L Wang and Z Shao ldquoCyclin E expression and prognosis inbreast cancer patients a meta-analysis of published studiesrdquoCancer Investigation vol 24 no 6 pp 581ndash587 2006

[73] Y J Zhou Y T Xie J Gu L Yan G X Guan and X Liu ldquoOver-expression of cyclin e isoforms correlates with poor prognosisin rectal cancerrdquo European Journal of Surgical Oncology vol 77no 12 pp 1078ndash1084 2011

[74] D M Bortner and M P Rosenberg ldquoInduction of mammarygland hyperplasia and carcinomas in transgenic mice express-ing human cyclin ErdquoMolecular and Cellular Biology vol 17 no1 pp 453ndash459 1997

[75] P L Porter K E Malone P J Heagerty et al ldquoExpressionof cell-cycle regulators p27(Kip1) and cyclin E alone and incombination correlate with survival in young breast cancerpatientsrdquo Nature Medicine vol 3 no 2 pp 222ndash225 1997

[76] Y Akama W Yasui H Yokozaki et al ldquoFrequent amplificationof the cyclin E gene in human gastric carcinomasrdquo JapaneseJournal of Cancer Research vol 86 no 7 pp 617ndash621 1995


[77] H K Kim I Park D S Heo et al ldquoCyclin E overexpression asan independent risk factor of visceral relapse in breast cancerrdquoEuropean Journal of Surgical Oncology vol 27 no 5 pp 464ndash471 2001

[78] J Tsihlias L Kapusta and J Slingerland ldquoThe prognosticsignificance of altered cyclin-dependent kinase inhibitors inhuman cancerrdquoAnnual Review ofMedicine vol 50 pp 401ndash4231999

[79] O Straume L Sviland and L A Akslen ldquoLoss of nuclear p16protein expression correlates with increased tumor cell prolif-eration (Ki-67) and poor prognosis in patients with verticalgrowth phase melanomardquo Clinical Cancer Research vol 6 no5 pp 1845ndash1853 2000

[80] H Zhang and X F Sun ldquoLoss of p27 expression predicts poorprognosis in patients with Dukesrsquo B stage or proximal colorectalcancerrdquo International Journal of Oncology vol 19 no 1 pp 49ndash52 2001

[81] A Juuti S Nordling J Louhimo J Lundin K Von Boguslaw-ski and C Haglund ldquoLoss of p27 expression is associated withpoor prognosis in stage I-II pancreatic cancerrdquo Oncology vol65 no 4 pp 371ndash377 2003

[82] T Tenjo M Toyoda J Okuda et al ldquoPrognostic significance ofp27(kip1) protein expression and spontaneous apoptosis inpatients with colorectal adenocarcinomasrdquo Oncology vol 58no 1 pp 45ndash51 2000

[83] Y Hu M Liao J Ding J Zhou and K Xu ldquoCo-deletion ofboth p15p16 genes correlates with poor prognosis non-smallcell lung cnacerrdquo Chinese Journal of Cancer Research vol 14 no3 pp 216ndash219 2002

[84] Y Matsuda T Ichida T Genda S Yamagiwa Y Aoyagi and HAsakura ldquoLoss of p16 contributes to p27 sequestration by cyclinD l-cyclin-dependent kinase 4 complexes and poor prognosis inhepatocellular carcinomardquo Clinical Cancer Research vol 9 no9 pp 3389ndash3396 2003

[85] S Fujiwara T Noguchi S Takeno Y Kimura S Fumoto andKKawahara ldquoHypermethylation of p16 gene promoter correlateswith loss of p16 expression that results in poorer prognosis inesophageal squamous cell carcinomasrdquo Diseases of the Esopha-gus vol 21 no 2 pp 125ndash131 2008

[86] U R Kees P R Burton C Lu and D L Baker ldquoHomozygousdeletion of the p16MTS1 gene in pediatric acute lymphoblasticleukemia is associated with unfavorable clinical outcomerdquoBlood vol 89 no 11 pp 4161ndash4166 1997

[87] Y Yamada Y Hatta K Murata et al ldquoDeletions of p15 andorp16 genes as a poor-prognosis factor in adult T-cell leukemiardquoJournal of Clinical Oncology vol 15 no 5 pp 1778ndash1785 1997

[88] H Tien J Tang W Tsay et al ldquoMethylation of the p15INK4Bgene in myelodysplastic syndrome it can be detected early atdiagnosis or during disease progression and is highly associatedwith leukaemic transformationrdquoBritish Journal ofHaematologyvol 112 no 1 pp 148ndash154 2001

[89] J Tsihlias L Kapusta and J Slingerland ldquoTheprognostic signif-icance of altered cyclin-dependent kinase inhibitors in humancancerrdquo Annual Review of Medicine vol 50 pp 401ndash423 1999

[90] S A Wander D Zhao and J M Slingerland ldquop27 a barometerof signaling deregulation and potential predictor of response totargeted therapiesrdquo Clinical Cancer Research vol 17 no 1 pp12ndash18 2011

[91] Y Takano Y Kato P J van Diest M Masuda H Mitomi andI Okayasu ldquoCyclin D2 overexpression and lack of p27 correlatepositively and cyclin E inversely with a poor prognosis in gastric

cancer casesrdquo American Journal of Pathology vol 156 no 2 pp585ndash594 2000

[92] A Pertia D Nikoleishvili O Trsintsadze N Gogokhia LManagadze and A Chkhotua ldquoLoss of p27(Kip1) CDKI is apredictor of poor recurrence-free and cancer-specific sur-vival in patients with renal cancerrdquo International Urology andNephrology vol 39 no 2 pp 381ndash387 2007

[93] T Abbas and A Dutta ldquoP21 in cancer intricate networks andmultiple activitiesrdquo Nature Reviews Cancer vol 9 no 6 pp400ndash414 2009

[94] S Giglio R Cirombella R Amodeo et al ldquoMicroRNA miR-24 promotes cell proliferation by targeting the CDKs inhibitorsp27Kip1 and p16INK4ardquo Journal of Cellular Physiology vol 228no 10 pp 2015ndash2023 2013

[95] I Ivanovska A S Ball R L Diaz et al ldquoMicroRNAs in themiR-106b family regulate p21CDKN1A and promote cell cycleprogressionrdquo Molecular and Cellular Biology vol 28 no 7 pp2167ndash2174 2008

[96] F Petrocca R Visone M R Onelli et al ldquoE2F1-regulatedmicroRNAs impair TGFbeta-dependent cell-cycle arrest andapoptosis in gastric cancerrdquo Cancer Cell vol 13 no 3 pp 272ndash286 2008

[97] S Wu S Huang J Ding et al ldquoMultiple microRNAs modulatep21Cip1Waf1 expression by directly targeting its 31015840 untranslatedregionrdquo Oncogene vol 29 no 15 pp 2302ndash2308 2010

[98] J Koh T Kubota T Koyama et al ldquoCombined antitumoractivity of 7-hydroxystaurosporine (UCN-01) and tamoxifenagainst human breast carcinoma in vitro and in vivordquo BreastCancer vol 10 no 3 pp 260ndash267 2002

[99] Y Kasuya Y Hosaka H Matsushima T Goto T Kitamuraand A Okuyama ldquoProminent induction of cyclin B1 in G2Mrenal cancer cells with butyrolactone 1rdquo International Journal ofUrology vol 10 no 6 pp 323ndash331 2003

[100] XM Liao andK N Leung ldquoIndirubin-31015840-oxime inducesmito-chondrial dysfunction and triggers growth inhibition and cellcycle arrest in human neuroblastoma cellsrdquo Oncology Reportsvol 29 no 1 pp 371ndash379 2013

[101] S Kim S J Choi Y Kim and H Kuh ldquoAnti-tumor activity ofnoble indirubin derivatives in human solid tumor models invitrordquo Archives of Pharmacal Research vol 32 no 6 pp 915ndash922 2009

[102] S Lapenna and A Giordano ldquoCell cycle kinases as therapeutictargets for cancerrdquo Nature Reviews Drug Discovery vol 8 no 7pp 547ndash566 2009

[103] M Kaliszczak H Patel S H Kroll et al ldquoDevelopment of acyclin-dependent kinase inhibitor devoid of ABC transporter-dependent drug resistancerdquo British Journal of Cancer vol 109no 9 pp 2356ndash2367 2013

[104] S C Chiu S Y Huang S P Chen et al ldquoTanshinone IIAinhibits human prostate cancer cells growth by induction ofendoplasmic reticulum stress in vitro and in vivordquo ProstateCancer and Prostatic Diseases vol 16 no 4 pp 315ndash322 2013

[105] L Wang G Wang D Yang et al ldquoEuphol arrests breast cancercells at the G1 phase through the modulation of cyclin D1 p21and p27 expressionrdquo Molecular Medicine Reports vol 8 no 4pp 1279ndash1285 2013

[106] C Haider M Grubinger E Reznıckova et al ldquoNovel inhibitorsof cyclin-dependent kinases combat hepatocellular carcinomawithout inducing chemoresistancerdquo Molecular Cancer Thera-peutics vol 12 no 10 pp 1947ndash1957 2013


[107] L C Ooi N Watanabe Y Futamura et al ldquoIdentification ofsmall molecule inhibitors of p27Kip1 ubiquitination by high-throughput screeningrdquo Cancer Science 2013

[108] T Gurcky R Jorda M Zatloukal et al ldquoA novel series of highlypotent 2 6 9-trisubstituted purine cyclin-dependent kinaseinhibitorsrdquo Journal of Medicinal Chemistry vol 56 no 15 pp6234ndash6247 2013

[109] A S Hamid J Li Y Wang et al ldquoRecombinant human decorinupregulates p57KIP2 expression in HepG2 hepatoma cell linesrdquoMolecular Medicine Reports vol 8 no 2 pp 511ndash516 2013

[110] P Bose G L Simmons and S Grant ldquoCyclin-dependent kinaseinhibitor therapy for hematologic malignanciesrdquo Expert Opin-ion on Investigational Drugs vol 22 no 6 pp 723ndash738 2013

[111] J L Dean CThangavel A KMcClendon C A Reed and E SKnudsen ldquoTherapeutic CDK46 inhibition in breast cancer keymechanisms of response and failurerdquo Oncogene vol 29 no 28pp 4018ndash4032 2010

[112] B Peng J Cao S Yi et al ldquoInhibition of proliferation andinduction of G1-phase cell-cycle arrest by dFMGEN a novelgenistein derivative in lung carcinoma A549 cellsrdquo Drug andChemical Toxicology vol 26 no 2 pp 196ndash294 2013

[113] T S Nagaria J LWilliams C Leduc et al ldquoFlavopiridol syner-gizes with sorafenib to induce cytotoxicity and potentiate anti-tumorigenic activity in EGFRHER-2 and mutant RASRAFbreast cancer model systemsrdquo Neoplasia vol 15 no 8 pp 939ndash951 2013

[114] M Kitagawa K Kitagawa Y Kotabe et al ldquoCell cycle regulationby long non-coding RNAsrdquoCellular andMolecular Life Sciencesvol 70 no 24 pp 4785ndash4794 2013

[115] A W Tong and J Nemunaitis ldquoModulation of miRNA activityin human cancer a new paradigm for cancer gene therapyrdquoCancer Gene Therapy vol 15 no 6 pp 341ndash355 2008

[116] M F Segura H S Greenwald D Hanniford et al ldquoMicroRNAand cutaneous melanoma from discovery to prognosis andtherapyrdquo Carcinogenesis vol 33 no 10 pp 1823ndash1832 2012

[117] BHinz andK Brune ldquoAntipyretic analgesics nonsteroidal anti-inflammatory drugs selective COX-2 inhibitors paracetamoland pyrazolinonesrdquo Handbook of Experimental Pharmacologyvol 177 pp 65ndash93 2007

[118] E R Kokoska G S Smith A B Wolff Y Deshpande and TAMiller ldquoNonsteroidal anti-inflammatory drugs attenuate epi-dermal growth factor- induced proliferation independent ofprostaglandin synthesis inhibitionrdquo Journal of Surgical Researchvol 84 no 2 pp 186ndash192 1999

[119] F M Giardiello ldquoNSAID-induced polyp regression in familialadenomatous polyposis patientsrdquo Gastroenterology Clinics ofNorth America vol 25 no 2 pp 349ndash362 1996

[120] C J Barnes I L Cameron W E Hardman and M Lee ldquoNon-steroidol anti-inflammatory drug effect on crypt cell prolifera-tion and apoptosis during initiation of rat colon carcinogenesisrdquoBritish Journal of Cancer vol 77 no 4 pp 573ndash580 1998

[121] E Giovannucci K M Egan D J Hunter et al ldquoAspirin and therisk of colorectal cancer in womenrdquoTheNew England Journal ofMedicine vol 333 no 10 pp 609ndash614 1995

[122] D J A deGroot EG E deVriesH JMGroen and S de JongldquoNon-steroidal anti-inflammatory drugs to potentiate chem-otherapy effects from lab to clinicrdquo Critical Reviews in Oncol-ogyHematology vol 61 no 1 pp 52ndash69 2007

[123] A Janssen T J Maier S Schiffmann et al ldquoEvidence of COX-2 independent induction of apoptosis and cell cycle block inhuman colon carcinoma cells after S- or R-ibuprofen treatmentrdquo

European Journal of Pharmacology vol 540 no 1ndash3 pp 24ndash332006

[124] H Zhou L Huang Y Sun and B Rigas ldquoNitric oxide-donatingaspirin inhibits the growth of pancreatic cancer cells throughredox-dependent signalingrdquo Cancer Letters vol 273 no 2 pp292ndash299 2009

[125] T J Maier K Schilling R Schmidt G Geisslinger and SGrosch ldquoCyclooxygenase-2 (COX-2)-dependent and -inde-pendent anticarcinogenic effects of celecoxib in human coloncarcinoma cellsrdquo Biochemical Pharmacology vol 67 no 8 pp1469ndash1478 2004

[126] B A Narayanan M S Condon M C Bosland N KNarayanan and B S Reddy ldquoSuppression of N-methyl-N-nitrosoureatestosterone-induced rat prostate cancer growth bycelecoxib effects on cyclooxygenase-2 cell cycle regulation andapoptosis mechanism(s)rdquo Clinical Cancer Research vol 9 no 9pp 3503ndash3513 2003

[127] Y Huang L Chuang and W Hung ldquoMechanisms underlyingnonsteroidal anti-inflammatory drug-induced p27Kip1 expres-sionrdquo Molecular Pharmacology vol 62 no 6 pp 1515ndash15212002

[128] S J Shiff L Qiao L-L Tsai and B Rigas ldquoSulindac sulfide anaspirin-like compound inhibits proliferation causes cell cyclequiescence and induces apoptosis in HT-29 colon adenocarci-noma cellsrdquo Journal of Clinical Investigation vol 96 no 1 pp491ndash503 1995

[129] P Bonelli F M Tuccillo R Calemma et al ldquoChanges in thegene expression profile of gastric cancer cells in response toibuprofen a gene pathway analysisrdquoPharmacogenomics Journalvol 11 no 6 pp 412ndash418 2011

[130] Y Takada A Bhardwaj P Potdar and B B Aggarwal ldquoNon-steroidal anti-inflammatory agents differ in their ability tosuppress NF-120581B activation inhibition of expression of cycloox-ygenase-2 and cyclin D1 and abrogation of tumor cell prolifer-ationrdquo Oncogene vol 23 no 57 pp 9247ndash9258 2004

[131] D Thurnher M Bakroeva M Formanek B Knerer and JKornfehl ldquoNon-steroidal anti-inflammatory drugs inhibittelomerase activity in head and neck squamous carcinoma celllinesrdquo Head and Neck vol 23 no 12 pp 1049ndash1055 2001

[132] H Sawaoka S Kawano S Tsuji et al ldquoCyclooxygenase-2inhibitors suppress the growth of gastric cancer xenografts viainduction of apoptosis in nude micerdquo American Journal ofPhysiologymdashGastrointestinal and Liver Physiology vol 274 no6 pp G1061ndashG1067 1998

[133] M J Aryankalayil S T Palayoor D Cerna M T Falduto SR Magnuson and C N Coleman ldquoNS-398 ibuprofen andcyclooxygenase-2 RNA interference produce significantly dif-ferent gene expression profiles in prostate cancer cellsrdquo Molec-ular Cancer Therapeutics vol 8 no 1 pp 261ndash273 2009

[134] M Redpath CMGMarques C Dibden AWaddon R Lallaand S MacNeil ldquoIbuprofen and hydrogel-released ibuprofen inthe reduction of inflammation-inducedmigration inmelanomacellsrdquo British Journal of Dermatology vol 161 no 1 pp 25ndash332009

[135] E J Quann F Khwaja and D Djakiew ldquoThe p38 MAPK path-way mediates aryl propionic acid-induced messenger RNA sta-bility of p75NTR in prostate cancer cellsrdquo Cancer Research vol67 no 23 pp 11402ndash11410 2007

[136] J Andrews D Djakiew S Krygier and P Andrews ldquoSuperioreffectiveness of ibuprofen compared with other NSAIDs forreducing the survival of human prostate cancer cellsrdquo Cancer


Chemotherapy and Pharmacology vol 50 no 4 pp 277ndash2842002

[137] I Tegeder J Pfeilschifter and G Geisslinger ldquoCyclooxygenase-independent actions of cyclooxygenase inhibitorsrdquo The FASEBJournal vol 15 no 12 pp 2057ndash2072 2001

[138] J L Masferrer K M Leahy A T Koki et al ldquoAntiangiogenicand antitumor activities of cyclooxygenase-2 inhibitorsrdquoCancerResearch vol 60 no 5 pp 1306ndash1311 2000

[139] S V Ekholm and S I Reed ldquoRegulation of G1 cyclin-dependentkinases in the mammalian cell cyclerdquo Current Opinion in CellBiology vol 12 no 6 pp 676ndash684 2000

[140] J F Viallard F Lacombe F Belloc J L Pellegrin and J ReiffersldquoMolecular mechanisms controlling the cell cycle main consid-erations and implications in oncologyrdquo CancerRadiotherapievol 5 no 2 pp 109ndash129 2001

[141] K Kristjansdottir and J Rudolph ldquoCdc25 phosphatases andcancerrdquo Chemistry and Biology vol 11 no 8 pp 1043ndash10512004

[142] T Waldman K W Kinzler and B Vogelstein ldquop21 Is necessaryfor the p53-mediated G1 arrest in human cancer cellsrdquo CancerResearch vol 55 no 22 pp 5187ndash5190 1995

[143] H Cam and B D Dynlacht ldquoEmerging roles for E2F beyondtheG1S transition andDNA replicationrdquoCancer Cell vol 3 no4 pp 311ndash316 2003

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


in a precise sequential manner It is dictated by the accumu-lation of CDKCCN complexes and degradation of CCNsQuiescent cells in G0 enter the G1 phase in response toexternal stimuli such as mitogenic growth factors or on thebasis of internal needs Cyclin D (CCND) binds to andactivates CDK4 andor CDK6 depending on the cell typeThe complexCCNDCDK4 orCDK6 phosphorylates RbThecyclin E (CCNE) binds to and activates CDK2 resulting inphosphorylation of a different site on Rb The E2F transcrip-tion factors (EF1ndashEF5) dissociate from the hyperphosphoryl-ated form of Rb to activate the transcription of genes thatpromote S phase including the thymidylate synthase (TS) thedihydrofolate reductase (DHFR) and the DNA polymerase(POL) [11 12] From this point on the cell has passed theldquorestriction pointrdquo and becomes committed to the progres-sion in the cell cycle and independent of growth factorsCCNA and CCNE bind to CDK2 allowing the cell to pro-ceed through the S phase The complex CCNACDK2 facil-itates the transition from the S phase to the G2 phase Thecomplexes CCNBCDK1 accumulate in late G2 phase neces-sary for the progression of the cell through the M phase [13]Following completion of anaphase CCNB is degraded thusreturning the cell to G1 state which in the presence of stim-ulation by growth factors proceeds by successive cycles ofcell division CDC25 (A B and C) are also required for theactivation of CDK complexes that control progressionthrough the cell cycle Activation of CDKs can be achievedthrough dephosphorylation bymembers of the CDC25 phos-phatase family (CDC25A CDC25B andCDC25C) CDC25Aplays an important role at theG1S-phase transition CDC25Bundergoes activation during S phase and plays a role inactivating the mitotic kinase CDK1CCNB in the cytoplasmActive CDK1CCNB then phosphorylates and activatesCDC25C leading to a positive feedback mechanism and toentry into mitosis [14] The repair of DNA damage or apop-tosis occurs prevalently at checkpoint G1 while the integrityof the synthesizedDNA is examined atG2 to ensure the fidel-ity of the replicated genome [15 16]TheCDKs play an impor-tant role in the regulation of these checkpoints For examplein response to various stress signals TP53 a transcriptionfactor is activated and causes the transcriptional inductionof CDKN1A and the cell cycle arrest at the G1 checkpoint[10] The length of the individual phases of the cell cycle canvary depending on the cell type and the particular conditionsThe activity of CDKs during the cell cycle is controlled atmultiple levels including the association with CCN acti-vating transient expression and rapid degradation of CCNsposttranslational modifications by kinases and phosphatasesinteractions with CDKIs and intracellular translocations [9]

3 Cell Cycle in Cancer

Oncogenic alterations of CCNs CDKs CDKIs and othercomponents of Rb pathway have been reported in more than90 of human cancers [1 2 17ndash40] as summarized in Table 1

One of the most relevant alterations is represented bythe p16 (CDKN2A)-CCNDsCDK-Rb pathway frequentlyaltered in various types of cancers [41ndash43] In solid tumors

DNA synthesis (S)

Cyclin BCDK1

Cyclin ACDK1

Cyclin DCDK4 CDK6

Cyclin ECDK2

Cyclin ACDK2

G2 G1

Mitosis (M)

Mammaliancell cycle

Figure 1 The mammalian cell cycle Cyclins and cyclin-dependentkinases are referred to their specific phase of the cell cycle Followingmitogenic signals that promote the cell entry in the G1 phasethe progression through the cell cycle is regulated by sequentialactivation of cyclins and cyclin-dependent kinases

there is a correlation between genetic alterations within thispathway and clinical outcome in cancer patients [44] Anom-alies of CDKs and CCNDs are very frequent in the G1 phaseSeveral molecular mechanisms may be responsible for thesealterations including gene overexpression chromosomaltranslocations pointmutations insertions and deletionsmis-sense and frame shift mutation splicing or methylationCDKs are found overexpressed in several types of tumorsincluding sarcoma colon carcinoma and lymphoma [45ndash47] CDK overexpression can be caused by gene amplification[48] chromosome translocation or point mutations [49]that impair the kinase interaction with CDK inhibitors asthe case for CDK4 in some melanoma patients [50] CDKactivity can also be dysregulated by overexpression of thecyclin partner or inactivation of CDK inhibitors both eventsbeing quite common in tumors [51 52]The lack of regulationof CDKs by its inappropriate activation is essential for main-taining the malignant transformation Changes of CCND1gene expression have been reported in several neoplasias Inparticular CCND1 gene is induced (transactivation) by var-ious oncogenic signals including the activating mutationof ras genes src and mitogen-activated protein kinases(MAPK) [53 54] as well as myc [55 56] Moreover chromo-somal aberrations involving CCND1 have been reported inB-lymphocytic malignancy and multiple myeloma [57 58]CCND1 overexpression played a role in the pathogenesis ofmammary cancer in transgenic mice [59 60] and lymphoma[61]The dysregulation of CCNE is associated with hyperpro-liferation and malignant transformation [26] Overexpres-sion of CCNE1 has been linked to endometrial hyperplasiaandor carcinoma [25] CCNE1 is overexpressed in manyhuman tumors in particular breast cancer and also nonsmall





































Table3Fu

nctio

nalgroup

ingof

genesu

p-anddo

wnregulated

after

ibup

rofentre

atment

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

MCellcyclecheckp

oint

G1S

DNAdamage

checkp

oint

CDC2

5ANM

001789

mdashmdash

minus2090

mdashmdash

mdashminus240

8mdash

mdashCelld

ivision

cycle

25ho

molog

A(Spom

be)

CDC2

5BNM

021873

mdashmdash

mdashmdash

mdashmdash


Celld

ivision

cycle

25ho

molog

B(Spom

be)

CDC2

5CNM

001790

mdashmdash

minus2155minus2306


mdashmdash

Celld

ivision

cycle

25ho

molog

C(Spom

be)

p53-depend

entG

1SDNADam

age

checkp

oint

TP53

NM

000546

mdash244

72503

mdashmdash

mdash3150

mdash3650

Tumor

proteinp53

CDKN

1ANM

078467

mdash3074

2789

mdashmdash

mdashmdash

minus2254minus2203

Cyclin-depend

entk

inase

inhibitor1A(p21C

ip1)

GADD45A

NM

001924

mdashmdash

mdashmdash

mdashmdash

4194

2493

2047

Growth

arrestand

DNA-

damage-indu

ciblealph

a

GADD45B

NM

015675

2352

mdash2191

2864

2792

2899

4016

5479

5200

Growth

arrestand

DNA-

damage-indu

ciblebeta

GADD45G

NM

006705

mdash2208

2661

2379

4796

5081

1412

12253

2Growth

arrestand

DNA-

damage-indu

ciblegamma

TP53IN

P1NM

033285

2033

3475

5494

3071

mdashmdash

mdashmdash

mdashTu

mor

protein53

indu

cible

nucle

arprotein1

TP53I3

NM

004881

mdashmdash

mdashmdash

mdash2214

mdashmdash

240

6Tu

mor

proteinp53indu

cible

protein3

MDM2

NM

002392

2534

mdash2868

3019

4111

3165

mdash4369

mdashMdm

2p53bind

ingprotein

homolog

(mou

se)


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

MCellcyclemito

ticG1S

transition

CCNE1

NM

001238

mdashmdash

mdashmdash


mdashmdash

CyclinE1

CDKN

1ANM

078467

mdash3074

2789

mdashmdash

mdashmdash

minus2254minus2203

Cyclin-depend

entk

inase

inhibitor1A(p21C

ip1)

CDKN

1BNM

0040

64mdash

mdashmdash

mdashmdash

2477

2432

3367

4938

Cyclin-depend

entk

inase

inhibitor1B(p27K

ip1)

CDC2

NM

001786

mdashmdash


Celld

ivision

cellcycle

2RB

1NM

000321

mdashmdash

mdashmdash

mdashmdash

mdash2872

2473

Retin

oblasto

ma1

DNAreplicationE

2Fmediatedregu

latio

nof

DNAreplication

E2F1

NM

005225

mdashmdash



E2Ftranscrip

tionfactor

1

CDT1

NM

030928

mdashmdash


Chromatin

licensin

gandDNA

replicationfactor

1PC

NA

NM

002592

mdashmdash


Proliferatin

gcellnu

clear

antig

enDHFR

NM

000791

mdashmdash

mdashmdash

minus246


12Dihydrofolateredu

ctase

TYMS

NM

001071

mdashmdash



76minus11282

Thym

idylates

ynthase

POLA

2NM

002689

mdashmdash

mdashmdash

mdashmdash


DNApo

lymerasea

lpha2

POLA

1NM

016937

mdashmdash

mdashmdash

mdashmdash

minus2098

mdashminus2014

DNApo

lymerasea

lpha

1


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

POLD

2NM

006230

mdashmdash

mdashmdash


DNApo

lymerase

delta

2

POLD

3NM

006591

mdashmdash

minus2244

mdashminus2135

mdashminus2992

mdashmdash

DNApo

lymerase

delta

3PO

LE2

NM

002692

mdashmdash


DNApo

lymerasee

psilo

n2

POLE

4NM

019896

mdashmdash

mdashmdash

mdashminus2154

mdashminus2118

mdashDNApo

lymerasee

psilo

n4

ORC

1LNM

004153

mdashmdash


mdashOrig

inrecogn

ition

complex

subu

nit1-like

(yeast)

RRM2

NM

001034

mdashmdash

mdashminus246


Ribo

nucle

otider

eductase

M2

subu

nit

Regu

latio

nof

DNA

replication

ORC

2LNM

006190

mdashmdash

mdashmdash

mdashmdash

2288

2727

2570

Orig

inrecogn

ition

complex

subu

nit2-like

(yeast)

ORC

4LNM

002552

mdashmdash

mdashmdash

mdash3216

2171

mdashmdash

Orig

inrecogn

ition

complex

subu

nit4

-like

(yeast)

ORC

5LNM

181747

mdashmdash

mdashmdash

mdashmdash

2209

2902

2765

Orig

inrecogn

ition

complex

subu

nit5-like

(yeast)

ORC

6LNM

014321

mdashmdash

mdashminus2348

mdashmdash

mdashminus2415

mdashOrig

inrecogn

ition

complex

subu

nit6

-like

(yeast)

MCM

2NM

004526

mdashmdash

mdashminus266


Minichrom

osom

emaintenance

complex

compo

nent

2

MCM

3NM

002388

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

3

MCM

4NM

005914

mdashmdash


4minus3923minus504

0minus73

44Minichrom

osom

emaintenance

complex

compo

nent

4

MCM

5NM

006739

mdashmdash



Minichrom

osom

emaintenance

complex

compo

nent

5


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

MCM

6NM

005915

mdashmdash


mdashmdash

mdashmdash

Minichrom

osom

emaintenance

complex

compo

nent

6

MCM

7NM

182776

mdashmdash

minus2175minus2563

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

7

TK1

NM

003258

mdashmdash


6minus75

57minus24478minus75

67minus5991

Thym

idinek

inase1solub

le

FEN1

NM

004111

mdashmdash

minus2008minus2268

mdashminus2110


Flap

structure-specific

endo

nucle

ase1

LIG1

NM

000234

mdashmdash


4minus6958minus564

0DNALigase

1

Apop

tosis

CASP

3NM

004346

mdashmdash

mdashmdash

mdash2097

mdash2239

2112

Caspase-3

CASP

7NM

033339

mdashmdash

mdashmdash

2124

2097

2050

2335

2393

Caspase-7

CASP

8NM

033356

mdashmdash

mdashmdash

mdashmdash

3686

2420

3056

Caspase-8

CASP

9NM

001229

mdashmdash

mdashmdash

mdashmdash

mdash2221

2257

Caspase-9

CASP

10NM

032977

mdashmdash

mdashmdash

mdashmdash

mdash3617

4514

Caspase-10

DFFA

NM

213566

mdashmdash

mdashmdash

mdashmdash

mdashmdash

2034

DNAfragmentatio

nfactor

45kD

aalph

apolypeptid

e

DFF

BNM

001004

285

mdashmdash

mdashmdash

mdashmdash

4015

2121

4583

DNAfragmentatio

nfactor40

kDbetap

olypeptid

e(caspase-activated

DNase)






6 Conclusions


Disclosure




References






























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




















































Table3Fu

nctio

nalgroup

ingof

genesu

p-anddo

wnregulated

after

ibup

rofentre

atment

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

MCellcyclecheckp

oint

G1S

DNAdamage

checkp

oint

CDC2

5ANM

001789

mdashmdash

minus2090

mdashmdash

mdashminus240

8mdash

mdashCelld

ivision

cycle

25ho

molog

A(Spom

be)

CDC2

5BNM

021873

mdashmdash

mdashmdash

mdashmdash


Celld

ivision

cycle

25ho

molog

B(Spom

be)

CDC2

5CNM

001790

mdashmdash

minus2155minus2306


mdashmdash

Celld

ivision

cycle

25ho

molog

C(Spom

be)

p53-depend

entG

1SDNADam

age

checkp

oint

TP53

NM

000546

mdash244

72503

mdashmdash

mdash3150

mdash3650

Tumor

proteinp53

CDKN

1ANM

078467

mdash3074

2789

mdashmdash

mdashmdash

minus2254minus2203

Cyclin-depend

entk

inase

inhibitor1A(p21C

ip1)

GADD45A

NM

001924

mdashmdash

mdashmdash

mdashmdash

4194

2493

2047

Growth

arrestand

DNA-

damage-indu

ciblealph

a

GADD45B

NM

015675

2352

mdash2191

2864

2792

2899

4016

5479

5200

Growth

arrestand

DNA-

damage-indu

ciblebeta

GADD45G

NM

006705

mdash2208

2661

2379

4796

5081

1412

12253

2Growth

arrestand

DNA-

damage-indu

ciblegamma

TP53IN

P1NM

033285

2033

3475

5494

3071

mdashmdash

mdashmdash

mdashTu

mor

protein53

indu

cible

nucle

arprotein1

TP53I3

NM

004881

mdashmdash

mdashmdash

mdash2214

mdashmdash

240

6Tu

mor

proteinp53indu

cible

protein3

MDM2

NM

002392

2534

mdash2868

3019

4111

3165

mdash4369

mdashMdm

2p53bind

ingprotein

homolog

(mou

se)


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

MCellcyclemito

ticG1S

transition

CCNE1

NM

001238

mdashmdash

mdashmdash


mdashmdash

CyclinE1

CDKN

1ANM

078467

mdash3074

2789

mdashmdash

mdashmdash

minus2254minus2203

Cyclin-depend

entk

inase

inhibitor1A(p21C

ip1)

CDKN

1BNM

0040

64mdash

mdashmdash

mdashmdash

2477

2432

3367

4938

Cyclin-depend

entk

inase

inhibitor1B(p27K

ip1)

CDC2

NM

001786

mdashmdash


Celld

ivision

cellcycle

2RB

1NM

000321

mdashmdash

mdashmdash

mdashmdash

mdash2872

2473

Retin

oblasto

ma1

DNAreplicationE

2Fmediatedregu

latio

nof

DNAreplication

E2F1

NM

005225

mdashmdash



E2Ftranscrip

tionfactor

1

CDT1

NM

030928

mdashmdash


Chromatin

licensin

gandDNA

replicationfactor

1PC

NA

NM

002592

mdashmdash


Proliferatin

gcellnu

clear

antig

enDHFR

NM

000791

mdashmdash

mdashmdash

minus246


12Dihydrofolateredu

ctase

TYMS

NM

001071

mdashmdash



76minus11282

Thym

idylates

ynthase

POLA

2NM

002689

mdashmdash

mdashmdash

mdashmdash


DNApo

lymerasea

lpha2

POLA

1NM

016937

mdashmdash

mdashmdash

mdashmdash

minus2098

mdashminus2014

DNApo

lymerasea

lpha

1


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

POLD

2NM

006230

mdashmdash

mdashmdash


DNApo

lymerase

delta

2

POLD

3NM

006591

mdashmdash

minus2244

mdashminus2135

mdashminus2992

mdashmdash

DNApo

lymerase

delta

3PO

LE2

NM

002692

mdashmdash


DNApo

lymerasee

psilo

n2

POLE

4NM

019896

mdashmdash

mdashmdash

mdashminus2154

mdashminus2118

mdashDNApo

lymerasee

psilo

n4

ORC

1LNM

004153

mdashmdash


mdashOrig

inrecogn

ition

complex

subu

nit1-like

(yeast)

RRM2

NM

001034

mdashmdash

mdashminus246


Ribo

nucle

otider

eductase

M2

subu

nit

Regu

latio

nof

DNA

replication

ORC

2LNM

006190

mdashmdash

mdashmdash

mdashmdash

2288

2727

2570

Orig

inrecogn

ition

complex

subu

nit2-like

(yeast)

ORC

4LNM

002552

mdashmdash

mdashmdash

mdash3216

2171

mdashmdash

Orig

inrecogn

ition

complex

subu

nit4

-like

(yeast)

ORC

5LNM

181747

mdashmdash

mdashmdash

mdashmdash

2209

2902

2765

Orig

inrecogn

ition

complex

subu

nit5-like

(yeast)

ORC

6LNM

014321

mdashmdash

mdashminus2348

mdashmdash

mdashminus2415

mdashOrig

inrecogn

ition

complex

subu

nit6

-like

(yeast)

MCM

2NM

004526

mdashmdash

mdashminus266


Minichrom

osom

emaintenance

complex

compo

nent

2

MCM

3NM

002388

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

3

MCM

4NM

005914

mdashmdash


4minus3923minus504

0minus73

44Minichrom

osom

emaintenance

complex

compo

nent

4

MCM

5NM

006739

mdashmdash



Minichrom

osom

emaintenance

complex

compo

nent

5


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

MCM

6NM

005915

mdashmdash


mdashmdash

mdashmdash

Minichrom

osom

emaintenance

complex

compo

nent

6

MCM

7NM

182776

mdashmdash

minus2175minus2563

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

7

TK1

NM

003258

mdashmdash


6minus75

57minus24478minus75

67minus5991

Thym

idinek

inase1solub

le

FEN1

NM

004111

mdashmdash

minus2008minus2268

mdashminus2110


Flap

structure-specific

endo

nucle

ase1

LIG1

NM

000234

mdashmdash


4minus6958minus564

0DNALigase

1

Apop

tosis

CASP

3NM

004346

mdashmdash

mdashmdash

mdash2097

mdash2239

2112

Caspase-3

CASP

7NM

033339

mdashmdash

mdashmdash

2124

2097

2050

2335

2393

Caspase-7

CASP

8NM

033356

mdashmdash

mdashmdash

mdashmdash

3686

2420

3056

Caspase-8

CASP

9NM

001229

mdashmdash

mdashmdash

mdashmdash

mdash2221

2257

Caspase-9

CASP

10NM

032977

mdashmdash

mdashmdash

mdashmdash

mdash3617

4514

Caspase-10

DFFA

NM

213566

mdashmdash

mdashmdash

mdashmdash

mdashmdash

2034

DNAfragmentatio

nfactor

45kD

aalph

apolypeptid

e

DFF

BNM

001004

285

mdashmdash

mdashmdash

mdashmdash

4015

2121

4583

DNAfragmentatio

nfactor40

kDbetap

olypeptid

e(caspase-activated

DNase)






6 Conclusions


Disclosure




References






























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

MCellcyclemito

ticG1S

transition

CCNE1

NM

001238

mdashmdash

mdashmdash


mdashmdash

CyclinE1

CDKN

1ANM

078467

mdash3074

2789

mdashmdash

mdashmdash

minus2254minus2203

Cyclin-depend

entk

inase

inhibitor1A(p21C

ip1)

CDKN

1BNM

0040

64mdash

mdashmdash

mdashmdash

2477

2432

3367

4938

Cyclin-depend

entk

inase

inhibitor1B(p27K

ip1)

CDC2

NM

001786

mdashmdash


Celld

ivision

cellcycle

2RB

1NM

000321

mdashmdash

mdashmdash

mdashmdash

mdash2872

2473

Retin

oblasto

ma1

DNAreplicationE

2Fmediatedregu

latio

nof

DNAreplication

E2F1

NM

005225

mdashmdash



E2Ftranscrip

tionfactor

1

CDT1

NM

030928

mdashmdash


Chromatin

licensin

gandDNA

replicationfactor

1PC

NA

NM

002592

mdashmdash


Proliferatin

gcellnu

clear

antig

enDHFR

NM

000791

mdashmdash

mdashmdash

minus246


12Dihydrofolateredu

ctase

TYMS

NM

001071

mdashmdash



76minus11282

Thym

idylates

ynthase

POLA

2NM

002689

mdashmdash

mdashmdash

mdashmdash


DNApo

lymerasea

lpha2

POLA

1NM

016937

mdashmdash

mdashmdash

mdashmdash

minus2098

mdashminus2014

DNApo

lymerasea

lpha

1


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

POLD

2NM

006230

mdashmdash

mdashmdash


DNApo

lymerase

delta

2

POLD

3NM

006591

mdashmdash

minus2244

mdashminus2135

mdashminus2992

mdashmdash

DNApo

lymerase

delta

3PO

LE2

NM

002692

mdashmdash


DNApo

lymerasee

psilo

n2

POLE

4NM

019896

mdashmdash

mdashmdash

mdashminus2154

mdashminus2118

mdashDNApo

lymerasee

psilo

n4

ORC

1LNM

004153

mdashmdash


mdashOrig

inrecogn

ition

complex

subu

nit1-like

(yeast)

RRM2

NM

001034

mdashmdash

mdashminus246


Ribo

nucle

otider

eductase

M2

subu

nit

Regu

latio

nof

DNA

replication

ORC

2LNM

006190

mdashmdash

mdashmdash

mdashmdash

2288

2727

2570

Orig

inrecogn

ition

complex

subu

nit2-like

(yeast)

ORC

4LNM

002552

mdashmdash

mdashmdash

mdash3216

2171

mdashmdash

Orig

inrecogn

ition

complex

subu

nit4

-like

(yeast)

ORC

5LNM

181747

mdashmdash

mdashmdash

mdashmdash

2209

2902

2765

Orig

inrecogn

ition

complex

subu

nit5-like

(yeast)

ORC

6LNM

014321

mdashmdash

mdashminus2348

mdashmdash

mdashminus2415

mdashOrig

inrecogn

ition

complex

subu

nit6

-like

(yeast)

MCM

2NM

004526

mdashmdash

mdashminus266


Minichrom

osom

emaintenance

complex

compo

nent

2

MCM

3NM

002388

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

3

MCM

4NM

005914

mdashmdash


4minus3923minus504

0minus73

44Minichrom

osom

emaintenance

complex

compo

nent

4

MCM

5NM

006739

mdashmdash



Minichrom

osom

emaintenance

complex

compo

nent

5


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

MCM

6NM

005915

mdashmdash


mdashmdash

mdashmdash

Minichrom

osom

emaintenance

complex

compo

nent

6

MCM

7NM

182776

mdashmdash

minus2175minus2563

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

7

TK1

NM

003258

mdashmdash


6minus75

57minus24478minus75

67minus5991

Thym

idinek

inase1solub

le

FEN1

NM

004111

mdashmdash

minus2008minus2268

mdashminus2110


Flap

structure-specific

endo

nucle

ase1

LIG1

NM

000234

mdashmdash


4minus6958minus564

0DNALigase

1

Apop

tosis

CASP

3NM

004346

mdashmdash

mdashmdash

mdash2097

mdash2239

2112

Caspase-3

CASP

7NM

033339

mdashmdash

mdashmdash

2124

2097

2050

2335

2393

Caspase-7

CASP

8NM

033356

mdashmdash

mdashmdash

mdashmdash

3686

2420

3056

Caspase-8

CASP

9NM

001229

mdashmdash

mdashmdash

mdashmdash

mdash2221

2257

Caspase-9

CASP

10NM

032977

mdashmdash

mdashmdash

mdashmdash

mdash3617

4514

Caspase-10

DFFA

NM

213566

mdashmdash

mdashmdash

mdashmdash

mdashmdash

2034

DNAfragmentatio

nfactor

45kD

aalph

apolypeptid

e

DFF

BNM

001004

285

mdashmdash

mdashmdash

mdashmdash

4015

2121

4583

DNAfragmentatio

nfactor40

kDbetap

olypeptid

e(caspase-activated

DNase)






6 Conclusions


Disclosure




References






























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

POLD

2NM

006230

mdashmdash

mdashmdash


DNApo

lymerase

delta

2

POLD

3NM

006591

mdashmdash

minus2244

mdashminus2135

mdashminus2992

mdashmdash

DNApo

lymerase

delta

3PO

LE2

NM

002692

mdashmdash


DNApo

lymerasee

psilo

n2

POLE

4NM

019896

mdashmdash

mdashmdash

mdashminus2154

mdashminus2118

mdashDNApo

lymerasee

psilo

n4

ORC

1LNM

004153

mdashmdash


mdashOrig

inrecogn

ition

complex

subu

nit1-like

(yeast)

RRM2

NM

001034

mdashmdash

mdashminus246


Ribo

nucle

otider

eductase

M2

subu

nit

Regu

latio

nof

DNA

replication

ORC

2LNM

006190

mdashmdash

mdashmdash

mdashmdash

2288

2727

2570

Orig

inrecogn

ition

complex

subu

nit2-like

(yeast)

ORC

4LNM

002552

mdashmdash

mdashmdash

mdash3216

2171

mdashmdash

Orig

inrecogn

ition

complex

subu

nit4

-like

(yeast)

ORC

5LNM

181747

mdashmdash

mdashmdash

mdashmdash

2209

2902

2765

Orig

inrecogn

ition

complex

subu

nit5-like

(yeast)

ORC

6LNM

014321

mdashmdash

mdashminus2348

mdashmdash

mdashminus2415

mdashOrig

inrecogn

ition

complex

subu

nit6

-like

(yeast)

MCM

2NM

004526

mdashmdash

mdashminus266


Minichrom

osom

emaintenance

complex

compo

nent

2

MCM

3NM

002388

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

3

MCM

4NM

005914

mdashmdash


4minus3923minus504

0minus73

44Minichrom

osom

emaintenance

complex

compo

nent

4

MCM

5NM

006739

mdashmdash



Minichrom

osom

emaintenance

complex

compo

nent

5


Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

MCM

6NM

005915

mdashmdash


mdashmdash

mdashmdash

Minichrom

osom

emaintenance

complex

compo

nent

6

MCM

7NM

182776

mdashmdash

minus2175minus2563

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

7

TK1

NM

003258

mdashmdash


6minus75

57minus24478minus75

67minus5991

Thym

idinek

inase1solub

le

FEN1

NM

004111

mdashmdash

minus2008minus2268

mdashminus2110


Flap

structure-specific

endo

nucle

ase1

LIG1

NM

000234

mdashmdash


4minus6958minus564

0DNALigase

1

Apop

tosis

CASP

3NM

004346

mdashmdash

mdashmdash

mdash2097

mdash2239

2112

Caspase-3

CASP

7NM

033339

mdashmdash

mdashmdash

2124

2097

2050

2335

2393

Caspase-7

CASP

8NM

033356

mdashmdash

mdashmdash

mdashmdash

3686

2420

3056

Caspase-8

CASP

9NM

001229

mdashmdash

mdashmdash

mdashmdash

mdash2221

2257

Caspase-9

CASP

10NM

032977

mdashmdash

mdashmdash

mdashmdash

mdash3617

4514

Caspase-10

DFFA

NM

213566

mdashmdash

mdashmdash

mdashmdash

mdashmdash

2034

DNAfragmentatio

nfactor

45kD

aalph

apolypeptid

e

DFF

BNM

001004

285

mdashmdash

mdashmdash

mdashmdash

4015

2121

4583

DNAfragmentatio

nfactor40

kDbetap

olypeptid

e(caspase-activated

DNase)






6 Conclusions


Disclosure




References






























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table3Con

tinued

Symbo

lAc

cessno

Fo

ldchange

Descriptio

n24

h48

h72

h40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M40

0120583M

600120583

M800120583

M

MCM

6NM

005915

mdashmdash


mdashmdash

mdashmdash

Minichrom

osom

emaintenance

complex

compo

nent

6

MCM

7NM

182776

mdashmdash

minus2175minus2563

mdashmdash


Minichrom

osom

emaintenance

complex

compo

nent

7

TK1

NM

003258

mdashmdash


6minus75

57minus24478minus75

67minus5991

Thym

idinek

inase1solub

le

FEN1

NM

004111

mdashmdash

minus2008minus2268

mdashminus2110


Flap

structure-specific

endo

nucle

ase1

LIG1

NM

000234

mdashmdash


4minus6958minus564

0DNALigase

1

Apop

tosis

CASP

3NM

004346

mdashmdash

mdashmdash

mdash2097

mdash2239

2112

Caspase-3

CASP

7NM

033339

mdashmdash

mdashmdash

2124

2097

2050

2335

2393

Caspase-7

CASP

8NM

033356

mdashmdash

mdashmdash

mdashmdash

3686

2420

3056

Caspase-8

CASP

9NM

001229

mdashmdash

mdashmdash

mdashmdash

mdash2221

2257

Caspase-9

CASP

10NM

032977

mdashmdash

mdashmdash

mdashmdash

mdash3617

4514

Caspase-10

DFFA

NM

213566

mdashmdash

mdashmdash

mdashmdash

mdashmdash

2034

DNAfragmentatio

nfactor

45kD

aalph

apolypeptid

e

DFF

BNM

001004

285

mdashmdash

mdashmdash

mdashmdash

4015

2121

4583

DNAfragmentatio

nfactor40

kDbetap

olypeptid

e(caspase-activated

DNase)






6 Conclusions


Disclosure




References






























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















6 Conclusions


Disclosure




References






























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















production equipment for photovoltaics - singulus

Documents