Survival and apoptosisA dysregulated balance in colorectal cancer?

Isabel FabregatInstitut d’Invetigació Biomèdica de Bellvitge

L’Hospitalet, Barcelona, Spainifabregat@idibell.org

Proliferation Cell death

Homeostasis

PROLIFE -

RACIÓN

MUERTE

CELULARPROLIFERATION

CELLDEATH

Proliferation Cell death

Tumor

PROLIFE-RACIÓN

MUERTECELULAR

-

CELLDEATH

PROLIFERATION

Signalling pathways involved in physiological apoptosis

CHARACTERISTICSCHARACTERISTICS OFOF APOPTOSISAPOPTOSIS

Selective proteolysis: CASPASES

- 116- 85

PARP

PHAGOCYTOSIS

- Phosphatidylserine exposition

Anexina VFITC

Chromatin condensation and nuclear fragmentation

DNA (IP)C

ell n

umbe

r

<2n

Fragmentation of the genomicDNA

180 bp

End

onuc

leas

e

Appearance of blebs in the cell membrane

No inflammation

Apoptotic bodies

CASPASESCASPASES: : CYSTEINCYSTEIN ASPARTATEASPARTATE PROTEASESPROTEASES

ACTIVE CASPASE (TETRAMER)

XXXAsp

PROTEOLYSIS (1000 SUBSTRATES)

INACTIVE PRECURSOR

CATALYTIC CENTERQACR(or Q/G)G

Asp Asp

Caspase-3, Caspase-7

(Initiator)

Apaf-1

AIF

Caspase-8

Bid

t-Bid

Bax

X-IA

P

SMAC/DIABLO

Cytochrome C

INTR

INSI

CPA

THW

AYpr

oCas

pase

-8Bcl-2Mcl-1

APOPTOSIS

EXTR

INSI

CPA

THW

AY

Caspase-9

Apoptosome

DISC

FasTNFRTRAILR

Cell death receptors

Ligands

Cellular stress

EndoG

(Initiator)

(Executioner)

BCL-2 FAMILY

Anti-apoptotic

Proapoptotic

Multidomain

BH3-only

Bcl-2 Bcl-xLMcl-1 Bcl-wBfl-1/A1 Diva/BooBrag-1

Bax BakBok/Mtd Bcl-XSBcl-GL

Bad Bid Bnip1 Mil1/Bcl-ramboBim Puma Bmf Nix/Bnip3LBik/Blk Noxa/apr Bfk Nip1/Bnip1Hrk/DP5 Bnip3 Bpr Bnip3h

BH4BH3BH1BH2TM

Cell damage sensors

Regulation ofmultidomain pro- and

antiapoptoticmembers

Caspase-3, Caspase-7

(Initiator)

Apaf-1

AIF

Caspase-8

Bid

t-Bid

Bax

X-IA

P

SMAC/DIABLO

Cytochrome C

INTR

INSI

CPA

THW

AYpr

oCas

pase

-8Bcl-2Mcl-1

APOPTOSIS

EXTR

INSI

CPA

THW

AY

Caspase-9

Apoptosome

DISC

FasTNFRTRAIL

Cell death receptors

Ligands

Cellular stress

EndoG

(Initiator)

(Executioner)

SIGNALLING PATHWAYS INVOLVED IN EPITHELIAL CELL SURVIVAL

Extracellular

Cytoplasm

Nucleus

RTK

GFs

EGFRs

c-MET

PI3K

AKT/PKB

mTOR

P70 S6K

Translation of cell cycleregulatory proteins

Stat

Ras

Raf

MEK1/2

ERK

ERK

P27kip1, c-Myc, c-Fos, cyclinB.

PTEN

BadForkhead

ProGFs

PROLIFERATION

SURVIVAL

EGF/HB-EGF/TGFα/amphiregulin

HGF

TACE / ADAMs

Two major pathways playing essential roles in epithelial cell survival

bcl-2, mcl-1

CREB

P90 RSK

Fas , other pro-apoptotic factors

SOME CYTOKINES PLAY DUAL ROLES IN CONTROLLING CELL DEATH

Extracellular

Cytoplasm

Nucleus

RTK

GFsEGF/HB-EGF/TGFα/amphiregulin

HGF

EGFRs

c-MET

PI3K

AKT/PKB

mTOR

P70 S6K

Translation of cell cycleregulatory proteins

Stat

Ras

Raf

MEK1/2

ERK

ERK

P27kip1, c-Myc, c-Fos, cyclinB.

TNFR1

IKKKs

IKK

NFκB

P-IκB

NFκB

c-IAPs, XIAP, c-FLIP, Survivin, TRAF2, Bcl-XL, Mcl-1, MnSOD, FHC

TNFα

TRADD-TRAF2-RIP

ASK1

MKK4,7

JNK

c-Jun

PTEN

CREB

P90 RSK

BadForkhead

TRADD- TRAF2-RIP, FADD

TRAIL-R1

TRAIL FAS L

FAS

Caspase-8,-10c-FLIP

Caspase-3,-6,-7

APOPTOSIS

Bid

Cyt c

Bcl-2 BclXL

Casp-9Apaf-1

Apoptosome

TACE / ADAMs

ProGFs

BadBax

Smac/Diablo

IAPsAKT

PROLIFERATIONSURVIVAL

APOPTOSIS

SURVIVALbcl-2, mcl-1

ROSGlutathione

ΔΨm

cit-C

APOPTOSIS

Caspase-9 Caspases 3 and 8

NADPHoxCHXAntioxidants

PI-3K

SURVIVAL

TGF-β TGF-αHB-EGF

TACE

Fetal rat hepatocytesFaO rat hepatoma cells

Epithelial-mesenchymal transitionprocesses

EGFR

Murillo et al. Oncogene 2005Caja et al. Cell. Signal. 2007

Ser-threo Kinase

Epithelial to Mesenchymal transitions:In the crossroads to apoptosis

resistance and cell invasion

RT - 3 5 8 16 24 36 48

snail

E-cadherin

+ TGFβ (h)

alb

TGFβ also changes the expression of some genes involved in epithelial to mesenchymal transitions

Caja et al. Cell Signal 2007

1 day 3 daysControl

E-CAD

DAPI

Control TGFβ

ControlTGFβ

FaO

Phalloidin

The EMT confer resistance to apoptosis. Snail activates survival pathways

Vega et al.Genes & Dev 2004

Increase in migration, apoptosis resistance…

METASTATIC POTENTIAL

Dysregulation of the apoptosis/survival pathway in

colorectal cancer

APOPTOSIS

PTEN

FAS

Bax, Bak

p53

SURVIVAL

Survivin

Bcl-2

HGF

β-catenin

EGFR ligands

PI-3K/AKTApaf1TβRII

P53, a guardian of the genome and a policeman of oncogenes

Puma, Noxa

Fas

Survival signals

Inflammation, colorectal cancer and NF-κB

Watson AJM, Crit Rev Onc Hematol, 2006

Inflammation, colorectal cancer and COX-2

Wand D and DuBois RN. Gut 2006

APOPTOSIS

PTEN

FAS

Bax, Bak

p53

SURVIVAL

SurvivinBcl-2

HGF

β-catenin

EGFR ligands

PI-3K/AKTApaf1TβRII

NF-κB

COX-2

APOPTOSIS

PTEN

FAS

Bax, Bak

p53

SURVIVALHGF

Survivin

β-catenin

Bcl-2

EGFR ligands

PI-3K/AKT

Apaf1

The balance between survival and apoptosis is dysregulated in colorectal cancer

NF-κB

COX-2

……Although multiple variables undoubtedly contribute to clinical outcome for patients diagnosed with early-stage colorectal cancer, these preliminary findings indicate that cIAP2, Apaf1, TUCAN, and Bcl-2 deserve further evaluation as potential prognostic markers. If confirmed in prospective studies, such prognostic biomarkers may help to guide treatment decisions for colorectal cancer patients, particularly regarding use of adjuvant chemotherapy.

Clin Cancer Res 2005;11(15) August 1, 2005

BiologBiologíía Molecular II. Facultada Molecular II. Facultadde Farmacia, de Farmacia, UCMUCM

Departamento de BioquDepartamento de Bioquíímica ymica yBiologBiologíía Molecular II. Facultada Molecular II. Facultad

de Farmacia, de Farmacia, UCMUCM

Irene CarmonaGaelle del CastilloAránzazu Sánchez

Margarita FernándezCésar Roncero

Laia CajaMiguel Murillo Conrad Ortiz

Esther BertranPatricia SanchoIsabel Fabregat

IDIBELL-Institut de Investigació Biomèdica de

BellvitgeL´Hospitalet, Barcelona

Instituto de Neurociencias, CSIC-Univ. Miguel Hernández, Alicante

Sonia Vega Ángela M. Nieto

CNIC , Instituto Salud Carlos III, Madrid

Alberto Álvarez-Barrientos

University of Vienna, Austria

Wolfgang Mikulits lab

Natalia Lazarevich(University of Moscow, Russia)