187

TNF-a (EC,,: 2.7, 8.2, and 2.4 pM, respectively), although a series of

othercylokinesandgrowth factors didnol have thiseffecl.Cort& also

increased 13AR density (EC,,: 30 nM) and markedly porentiated the

effects of IL-la, IL-U, and TN&a. Neither IL-1 nor cortisol influ-

enced Lhe. proportion of cell surface vs internalized BAR. The IL-l-

induced increase in BAR density was half-maximal after 6 h, was

reversible at a similar rate, and was blocked by I pM of cyclohcximide.

The effect of IL- 1 on LIAR was specific, as the density of glucocorticold

receptors, measurement by 3H-dexamethasone binding, was reduced by

IL-I. Both conisol and IL-1 potentiated the isoprolerenol-induced

mcrease in CAMP accumulation. IL-1 Inhibited cell proliferation and

thymidine uptake, and mcreased the adherence of A549 cells to the

plasticcultureflask,asquantifiedbyacellderachmen~assay.Theeffect

of IL-1 on cell adherence was not inhibited by cycloheximide. Cortisol

decreased cell adherence and prevented Ihe IL-l-induced increase m

adherence. Theresults Indicate that multiplceffectsofIL-1 inacultured

tumor cell line involve different mechanisms, suggesting heterogeneity

of IL- 1 R and/or coupling of IL- 1 R 10 distinct, nuclear, and nonnuclear,

effector pathways.

A human monocyte growth factor produced by lung cancer cells

Okabe T, Yasukawa K. The Third Deparmznf of Internal Medicine, Facuify of Medicme. Universilj of Tokyo, llongn 7-3-l. Bunkyo-ku. Tokyo 113. Cancer Res 1990;50:3863-5.

Human lung cancer cell lint, T3M.30, has been shown io produce a

growth factor that stimulalcs problcratlon of peripheral blood mon-

wyles. In Ihe prescncc of this factor, human circuladng monocyles

were able Lo prolifcratc in wuo. Gel exclusion chromatography of Ihe

condrlioned medium rcvealcd a smgle peak of monocyte growth-pro-

motingactlvi~ya~anapparentmolccularwcigh~of 16,OOO.Thegrowth-

promoting activity was absorbed to an anion-exchange column, Mono

Q, and clutcd wlh a salt gradlcnc as a single peak of bioactivity at 300

mM NaCI. When the sample was applied to a Vydac C, column, a

rcversc-phase ixgh-pcrformaoce liquid chmma~ogrdphy column, a single

peak of activity was observed at a concemration of 76% acetonilrile in

0.1% Uifluoroacctic acid. The monocytc growth-promoting activity

was heat stable at 56°C. 11 was partmlly dcsuoyed by trypsin. The

activity was lost alLr trcatmcnt with 2.mcrcapmethanol.

Aneoropeptideantagonist that inhibits thegrowthofsmallcelllung

cancer in vitro

Wall PI, Rozengurl E. GrowrhRegularionLaboratory,lmperral Cancer Research Fund. Lmcoln’s Inn Fields, London WCZA 3PX. Cancer Rcs

1990;50:3968-73.

In Ihe search for novel antlprobfcrative agenE for small cell lung

cancer (SCLC), WC found the ncuropeptide anragomst [Arg6,D-

Trp’~‘,MePheR]subscance P(6-I I) to be effective in vitro. In “urine

Swiss 3T3 cells [Arg6,D-Trp7,9.MePheR]subsllnce P(6-11) was identi-

fled as a potcnt inhibitor of vasoprcssin-stimulated DNA synthesis

whrch also blocks [‘Hlvasopressm bmding to spwfic cell-surface

rcceplors. It was a less potent antagonist of gastrin-releasing pcptidc

and bradykinin in these cells but did not block Ihe effects of other

mllogens. In SCLC cell lines, [Arg”.D-Trp7~P,MePheR]subslance P(6-

I I) inhibltcd colony-formauon in soft agarosc and growth in hquid

cullurc in a dose-dependent manner. It also blocked rcccplor-mediated

Ca” mobibzatlon induced by vasopressm, bradykinin. cholecystokinin,

galanin, gasuin-releasing pepudc, and ncurownsm. We suggest that

broad-specuum neuropcptide antagonists can block multiple autocrinc

and paracrinc growth leaps m SCLC and could bc useful therapeutic

agents.

Cloned low m&static variants from human lung carcinoma metas- tases

Varki NM, Tseng A, Vu TP, Estcs LA. Deparlmenl ofMedicine, Cancer Cenrer. Universify of Califorma. San Diego, La Jolla. CA 92093.

Anticancer Res 1990;10:637.43.

ClonaI subpopulations of neoplasuc cells were derived. in soft agar,

from the spontaneously metastazrng variant (MV522) of a human lung

carcinoma cell line. The ability of these clones 10 spontaneously

mciastasiz from subcwmcous sites m athymic mlcc was then tested. A

variation m mctastatic ability was cxpecwd with tic derivation ofsomc

low mctascalic clones and some high mcliL\taUc ones. However, all of

the denvcd clones, although equally tumortgemc, were less metasIatic

than Ihe parental variant. Thcsc clonal ccl1 lines can now be used in a

syslemaw analysis of cvcms asssoclatcd with Ihc reversion 10 a Icss

malignant staw.

Retinoic acid and epidermal growth factor binding in retinoid-

mediated invasion suppressed human lung carcinoma cells

Fazely F, Lcdinko N. Dana-Farber Cancer Research Inslr~ule, 44 Binnq Swea, Bosmn, MA 021 IS. Amicanccr Rcs 1990;10:667-70.

The cffcct of rctinolc acid (RA)-mduccd suppression of m wtro

mvasivc abUy of A549 human lung carcinoma cells on cellular

bmdingofRAand cpidcrmalgrowh faclor(EGF) was mvcs~lgawd.RA

inhibitloo of cell invasive potcmlal was accompamcd by a slgmficam

increase m specific hrgh affmity ceilular rctmoic acid bindmg protein

(CRABP) Icvcl. An approxlmalcly 2.7.fold mcreasc in cytosolic CRABP

was found in RA-trcawd cells (450 fm/mg prolem compared Lo I67 fm/

mg comrol cell protein). In conwarl, ‘“51-EGFligand hmmg was slmdar

for corm01 and ucatcd cells.

Expression of CYPlAl gene in patients with long cancer: Evidence

for cigarette smoke-induced gene expression in normal long tissue

and for altered gene regulation in primary pulmonary carcinomas

McLcmorc TL, Adclbcrg S, IJU MC cL al. Program Deve~opmenr Research Group. Developmental Therapeulics Program. Division of Cancer 7’reuumxl. Na~uvzul Cancer In,wuie, Bahesda, MD. J Nat1

Cancer Inst 1990 82:1333-Y.

The major polycyclic ammaw hydrocarbon mduclblc-cytochrome

P4501Al gene (CYPIAI) is prcsumcd Lo be imporlanr in pulmonary

carcinogcncsls and loxlcology because its product, the cyuxhrome

P450lAl -dependent (CYPlAl-dcpcndcnl) monooxygenasc, transforms

selected xenoblotics (including polycyclic aromatic hydrocarbon pro-

carcmogcns in clgarcllc smoke) Lo potent carcmogcmc mctabohles.

CYPIAI messenger RNA (“RNA) cxprcss~on has not, howcvcr, been

previously demonstrated m human pulmonary IISFUC. This report dc-

fines CYPIAI gene cxprcssion m normal lung twuc and primary

pulmonary carcmoma wsuc oblaincd at lhoracolomy from 56 paticms

with lung cancer. When Northcm blor hybridwation analysts were

pcrformcd,l7ofl9(X9~)and~crooff~vc(O%~samplcvofnormallung

lissuc from actwc cigaretlc smokers and nonsmokers, rcspcchvcly.

cxprcsscdthcnormal2.&ktlobascCYPl Al “RNA. Inaddlrmn,atrmc-

dependent dccrcasc m cxprcssmn of the CYPIAI gene was noted m

normal lung tissue from indiwduals who wcrc lormcr smokers, with a

decreascmcxprcssionoccurringascarlyas2 wccksfollowlngccssauon

of cigarcue smoking. Expression bccamc undcrcctablc m all patients

who had slopped smokmg mom than 6 weeks prior Lo study. When

CYPI A 1 gcnc cxprcsslon was cvaluatcd in lung cancers, mRNA levels

were delcctablc m one of four (25%) tumors from nonsmokers; two of

24 (8%) tumors from former smokers; and sewn of 15 (47%) tumors

from cigarctlc smokers. In addition, an approximately IO-kilobasc

CYPIAI RNA spccles, whvzh was not delectable m normal lung tissue,

was obscrvcd in fwc of ten (50%) of the lung cancers that cxprcssed Ihe

CYPIAI gcnc. There was no pasmvc assoclauon bcwcen CYPl Al

cnpression and lung cancer hwologic cell type nor bctwccn CYPl Al

“RNA lcvcls m malchcd normal lung ussuc and tumor Llssuc from

pauents with lungcanccr. Thcscrcsullsdcmonslratcaposlclvcassocla-

lion bctwccn acwc clgarcw smoking and CYPI Al gene expression m

normal human lung tissue. Morcovcr, CYPIAI gcnc exprcsslon was

documcntcd m many pulmonary carcmomas, and altered rcgula~ion of Ihc gene was also obscrvcd m scvcral lung tumor?.

Short- and long-term synergistic effects of human tumor necrosis

factor and interferon-gamma on AS49 human lung cancer cells

maintained in three-dimensional culture

Beaupain R, Martyre MC. Laboraloire d’lmmunopharmacologie Ex- perimenrale. UPR-405 CNRS. 15 Rue de I’Ecole de Medecine. 75270 Paris Cedex 06. Anticancer Res 199O;lO: 1061-6.

We have studied the short- and long-term effects of human recombi- nant tumor necrosis factor (TNF) and TNF/recombinant human inter- feron-gamma (IFN-gamma) mixtures on AS49 human lung carcinoma cells maintained in organotypic culture. Continuous treatments with 2 x 10; 2 x 10z, 2 x 101 and 2 x I@ U/ml TNF or with mixtures of TNF/ IFN-a at 2 x IO2 and IO3 U/ml, respectively, were administered. Nodule growth, cell proliferation and cell survival were studied. On the 2nd day of treatment with TNF, only the highest dose (2 x 104 U/ml) diminished cell proliferation significantly, as measured by tritiated thymidine uptake into DNA. On the 10th day, only the lowest TNF dose (2 x 10 U/ ml) Induced no significant growth inhibition. Necrosis and nodule disintegration were apparent in the 2 x 106 U/ml-treated noduleswhere DNAsynthesisdecreased. In thiscase,usingagarcloningassays,nocell survival could be observed. Similar results could be obtained with TNF at low concentration (2 x lo* U/ml) in combination with INF-gamma (lo3 U/ml), showing a synergistic effect on inhibition of cell prolifera- tion. In the long-term experiments, with the lower TNF doses, in situ evidence of regrowth was observed (outgrowing zones in the nodules) on about tbe40tb day of treatment, and nodule recovery was confirmed by the resumption of DNA synthesis measured on the 50th day of treatment. Noregrowth, however,occurred when theIFN-gammflNF combination was used, and the nodules disintegrated completely on the 35th day of treatment without evidence of any cellular survival.

Superior antiproliferative effects mediated by interferon-a en- trapped in liposomes against a newly established human lung cancer cell line Shin DM, Fidler IJ, Bucana CD, Fan D, Hong WK, Killion JJ. Deparr- ment of Medical Oncology. University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030. J Biol Re- sponse Modif 1990:9:35S-60.

The purpose of this study was to characterize the antiproliferative activity of a recombinant interferon-a (IFN-a) against a newly estab- lished human adenocarcinoma cell line (DMS4C) and to determine whetherIFN-a enuappedinmultilamellarliposomes hadsuperioranti- tumor effects compared with free IFN-a. Treatment of DMS4C cells with 100 U/ml of free IFN-a resulted in 34% cytostasis. IFNa encap- sulated in phosphatidylcholine/phosphatidylserine multilamellar ves- icles produced growth inhibition of 67%, which was significantly greater than that produced by free IFNa or by control liposomes containing only medium combined with free IFN-a. Moreover, kinetic analysis revealed that to produce significant cytolysis, free IFN-a had to be incubated with target cells for at least 24 h, whereas IFN-a encapsulated into liposomes required only 30 min of exposure.

Rationale for the use of chemotherapy in non-small cell lung cancer Miller TP. Cancer Center Clinics, Arizona Cancer Center. 151s N CampbellAve, Tucson, AZ85724. Semin 0nco11990;17:Supp17:1 l-3.

The vast majority of patients have disseminated non-small cell lung cancer (NSCLC) at the time of diagnosis. Data from numerous studies clearly indicate that the disease is metastatic in asymptomatic patients whoappear to haveclinically resectable tumors. Adenocarcinomais the histologic subtype that is most frequently metastatic, and its incidence appears to be increasing in a manner relative to other tumors. Of all of tbe prognostic factors, performance status appears to be most directly related to response rates. In other words, the higher the performance status, the higher me response rates. Tumor burden has been found to have an effect not only on performance status, but also on response to chemotherapy. Therefore systemic chemotherapy is urged as adjuvant treatment early in the course of NSCLC when performance status is highest and tumor burden lowest.

K-Ras oncogene activation as a prognostic marker in adenocar- cinema of the lung Slebos RJC, Kibbelaar RE, Dalesio 0 et al. Division of Experimental Therapy. Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam. New Engl J Med 1990;323:561-5.

Background. The capability of activated oncogenes to induce malig- nant transformation of immortalized cells in vitro has suggested that they have a similar role in the pathogenesis of human tumors. We

previously found that activation of the K-ras oncogene by a point mutation in codon 12 occurs in about one third of human lung adenocar- cinemas. Methods. We studied the clinical importance of this onco- gene-activation in 69 patients with lung adenoctucinoma in whom complete resection of the tumor was possible. The polymcrasc chain reaction was used to amplify ras-specific sequences of DNA isolated from frozen or paraffin-embedded tumor samples. Ras point mutations were subsequently detected and classified with the use of mutation- specific oligonucleotide probes. Results. Nineteen of the tumors har- bored a point mutation in codon 12 of the K-ras oncogene. There was no association between the K-ras point mutation and the age at diagnosis, sex, or presence of previous or concurrent neoplasms. Tumors positive for K-ras point mutations tended to be smaller and less differentiated than those without mutations. The K-ras codon- point mutation was a strong (and unfavorable) prognostic factor: 12 of the 19 patients with K-raspoint-mutation-positive tumorsdieddurmg the follow-upperiod, as compared with 16 of the 50 patients with no mutation in the K-ras oncogene (P = 0.002). This difference in prognosis was also reflected in the duration of disease-free survival (P = 0.038) and in the number of deaths due to cancer (P < 0.001). Conclusions. The presence of K-ras point mutations defines a subgroup of patients with lung adenocar cinema in whom the prognosis is very poor and disease-free survival is not usually long despite radical resection and a small tumor load.

Rolycyclicaromatic hydrocarbon - DNA adducts in lung tissue from lung cancer patients Van Schoomn FJ, Hillebrand MJX, Van Leeuwen FE et al. Division of Chemical Carcinogen&s. Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam. Carcinogcnesis 1990,11:1677-81.

In an attempt to probe for polycycbc aromatic hydrocarbon (PAH)- DNA adducts in human subjects resulting from smoking (or other chronic environmental exposure), lung tissue and lung tumours were obtained from patients hospitalized for lung cancer. DNA was Isolated from the tissue samples and examined both in an ELISA using a polyclonal antibody against (*)trans-7,8-dihydroxy-anti-9,10-epoxy- 7,8,9,10-tetrahydro benzo]a]pyr ene (BPDE) - DNA as well as by the nuclease PI-mediated modification of the 32P-post-labelling technique. The ELISA results showed BPDE DNA antigenicity in lung DNA from 6 out of 21 patients, and adduct levels ranged from 2 to 134 adducts per 108 nucleotides. For all 21 patients, the autoradiographs of chromato- grams of 32P-postlabelled digests of DNA from non-tumorous lung tissue showed a strong diagonal radioactive zone (DRZ). This DRZ was generally absent in tumorous tissue. DNA samples that were posttive in the ELISA contained a dominant spot within the DRZ that co-chroma- tographed with the major BPDE-DNA adduct (BPDE-dG). The quanti- ties of the BPDE-dG spots ranged from 2.1 to 42 adducts in IO9 nucleotides. These values were lower than the levels found in the ELISA but correlated well with the ELISA results (Kendall W = 0.97; P= 0.00). The levels of the DRZ adducts ranged from 1.9 to 34 adducts in 108 nucleotidcs. Correlations between smoking and DNA adduct levels were poor because of the small number of current smokers (n = 13). However, smokers of filter cigarettes had significantly lower DNA adduct levels compared with smokers of cigarettes without a filter (P = 0.02 by Fischer’s exact test).

Restriction fragment length polymorphism analysis of the L-myc gene locus in a case-control study of lung cancer Tamai S, Sugimura H, Caporaso NE et al. Laboratory of Human Carcinogenesis, DCE. NCI, NIH, Bethesda, MD 20892. Int J Cancer 1990;46:41 l-5.

The L-myc DNA-restriction fragment length polymorphism, re- vealed by EcoRI, has been studied in both a lung cancer case-control framework and a cohort of 40 nondiseased unrelated individuals. No association was found between the L-myc allelic frequencies and disease status, tumor stage or lung cancer histology. A strong associa- lion was, however, observed between the L-myc allelic frequencies and ethnic origin (black or white) of the subjects. Among American whites die allelic distribution at tbe L-myc proto-oncogene locus was almost identical to chat previously reported for Japanese subjects. Among the American black population there was a significantly higher frequency