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ANDREWS t.. .1CE PRODUCTS CAPITOL HEIGHTS, MD (K)

EXTRA COPY REPORT - Of THE COUNCIL FOR TOBACCO RESEARCH-U.S.A., Inc. 1982

Organization and Policy T>te Careil for Tobacco Ra.rrA-US.A. lec. Is 1be spoe.oriag .peey of a progrne ol research isto Suewbr of lobsooo un ..d health. It 1. tlt" out- powtt7 ol r orpnl:ation fofrd ..r1r i. i!S• by Ios.ooo s.suf.ctursn, pow.n rd w.rdawmes. Reaatsl wpoat bss bem An.ldy t6tougY a pro. Rum ol p.N.i.•.1d wrpplaw..td by ooWstb for wssaci+ wit• ieuiwt{o.e .ed LborswrM.. TS. C.arcll Io..a q.aw s.y n,arcU fscility. TU Sckatibc Ad.isory Soad ts Z1s Cor.ci www rtgul.rly 1o c.duN. .pplk.tio.s for research sappoct, jidgift tfsn..olely es 1lt@ e.un of scisstiie nrog1t .ed nMr..o.. T1. C.oarcU .+.rds ns..rch pr.w b i.depadea seiestists who .r. .r wnd oo.pkN wAestiffic fr.edo~w is oosducU.g tleir studies. Ur.nteea .b.. .n reaporiWe for repaU.g or puE1Wi.g t6e'r iodisp i. tb..coepted aie.- UAe m.oset - tlrough mediul sed .cie.tibc jour..b .ed socklhs. f F I 1982 REPORT o/ THE COUWCII. FOR TOBACCO RFSEAR®-U.S.A., t.e. WnLuw D. Hosss . ~ Cb.irsn. ~ ..7. . ~ .f... ~..+' ~abf m cn ~ '111E COUN('11. FOR TOBACCO RFSFARCII-U.S.A., iac. 110 t:..i 591h Street, New York, N.Y. 10022 m . m ~ J W ~ r r

S/aEN"1'IF'll: AI)VISI)RY IN)ARI) to The Council for Tobacco Research-U.S.A., Inc. a. u( Ikcember 31, 1982 I.EON O. JACOBSON. M I1., Chairmat losryh Rrjrnstrin Pro f,•stur of Biolosical Stient rs ( rmrritus ) I'rojrssrw tr/ thr 1)r/NOtment of Medicine (enrcritus) Univetsity of Chicago C'hica8o, Illinois RICIIARI) ). BING, M [). llirre un u/ ErttrrLnrvqul ('ardiukrtly and St irnti/ic I)eveltrpmrnt Ilumrn8ton Medical Research Institute, Pasadena, California I'tn/ritt.r o/ INrdit inC (rmrritut) l)nmvenily of Southern California School of Medicine I.cn AnBeles, California ROSWI7.l. K. BOl1TW[:LL, Pn.[). l'rtr/rta» ot OncokoRv McArdle Laboratory for Cancer Research University of Wisconsin Madison, Wisconsin DRI/MMOND 11. BOWDEN, M.D. I'roJessor and flrad Department of Pathology University of Manitoba Ilealth Sciences Center Winnipeg. Canada MICIIAEL 1. BRENNAN, M.D. President and Medical Director Michi¢an Cancer Foundation Detroit. Michigan 1OSEPH D. FELDMAN, M.D. Mrm6er, Research Institute of Scripps Clinic Scripps Clinic and Research Foundation La )olla, California WILLIAM l[. OARDNER, Pu.D. E. K. Nt.nt Pro/essra of Anatomy (emeritus) Yak l lniversily School of Medicine New Ilaven, ('onnecticul I PETER M. IIOWLEY, M.D. Laboratory of Pathology National Cancer Institute Bethesda, Maryland IfENRY T. LYNCH. M.D. Professor and Chairman Ikpartment of Preventive Medicine and Public Ikalth Creighton University School of Medicine Omaha, Nebraska G. BARRY PIIiRCE:, M.l). Amrriuan Concrr So.lrty Cintrnnial Research Pro/estor LMivcnily tr1 Colorado Ileatth Sciences Center (knver, ('olorado GORDON II. SATO, PH.D. Pro/essor of Biolt>ty University of Calilornia, San Diego La lolla, California SIIELDON C. SOMMERS Scientific Dira•tor, The Council for Tobacco Research-U.S.A., Inc. Clinical Pro/essor of Parbolory College of Physicians & Surgeons of Columbia University New York. New York Sekatlie Srafi.( The C.w..eil SHELDON C. SOMMERS, M.D. Sc•ienti/ic• Director ROBERT C. IiOCKETT, Pu.D. Research Director DONA1.1) 11. FORD, Ptt.D. VINCENT F. I.ISANTI, D.M.D. Associate Research Dirrtvor ~ Associate Rrsiarch Director DAVID STONE. Pu.D. Associate Research t)irector

Abstracts of Reports Fol{owint are abstracts, approved by the wthors. of reports on new resean:h ackmwvled6ins support from The Council that have appeared in scientific journds since publication of the 1951 Report. The name of the grant recipient is in italics. The abstracts are grouped under these headings: 1. Cancer-Relaled Studies. 11. The Respiratory System. 111. Heart and Circulation. IV. Neuropharmacology and Physiology. V. Pharmac.ology and Biochemistry. VI. Immunology and Adsptive Mechanisms. VII. Epidemiology. 1. Cancer-Related Studies MECHANISM OF ACfION OF BEN?d)/a/PYRENE AND NICOTINE ON HORMONE PRODUCIlON BY RAT PITUITARY TUMOR CELLS Although hormones have been associated with ieduction and ptopessan of m- Foms in many e.perimental systenu,lhe role of hoaeones in the process of initi.tiw and pnog reuiat of carcinoge.esis is.w ckaly de8ned as yet. lw the pesesw ancnrpl w .ndersund the mechanism of .ctiom of benm(a)Pyreae (BaP). a cyclic aomsfie hydevcarbon. and that of sticaine. the lobaceo.lkabid, the effects of these .gents on ppvlacqin (PRL) and growth honnone (GN) synthesis by rat pituilary tunwr oeMs in cuNure (GN cell!-) were studied. Treatment of GN cells with.icaline (0.1 •300 pg/ap neither affected the growth aor significantly s/1etel the general p.nerw of hmmone podaction in these ceds. BaP Y coecentntlions Rrerer than SpRhnl imeversibly inhibited the growth of these cells. The subkth.l eoncentntions o/ B.P. which did aot affect either (1) cell growth. or (2) amino acid tnnspoal or (l) rotal protein synthesis or desradaion, did however inhibil specifically harmone synthesis by these cells. More interestingly. concentntiota of sicoti.e. wbich did no1 affect either cell po.nh or hoxmonc synthesis. modulaed both of,these eeRuhY processes is drc presence of BaP. A concentration dependent stimulation of microsomal BaP rronoo.ylienase activity was observed in nicotine or BaP treated cells. Tbe effects of :fiese substances on aimulalion of BaP monoosylleaase activity seems 1o be additive. Nicotine also en- Mnced the associMioa ot adioaclivily (pRsttmsbly 1711B.P metabolites) with DNA i. /'111B.P treated cells. (t is concluded pist nicotine by itself did na demonwae any cywaosic e/fect not in/M.ence bunnone synthesis in G/I cells. However. ftieoti.e stimulated B.P inoonosygenase activily and the inleranion of /M1/B.P metabd'Nn witA ceHular tN1 A and also modulaled BaP induced inhibition o/ hormone synthesis is (;11 cells. Chakrabarli, S.. Nancs, S. D. .nd Aiiwns. D. K. SiocArnrira! md SfopAysiea/ Reseomh C-rirawns 101112):596-607, 1982. PFrom the I.baaory of Pfiarmacoloty. Harvard School of Dental Medicine and De• p.rlment of Ph.rmaolo6y. Ilarvan) Medical School. Boshin. 7

EFFECTS OF SELENIUM AND SULFUR ON METABOUSM OF BFJdZA>foIPYRENE BY HUMAN PULMONARY ALVEOLAR MAt.'ROPIIAGES The interactinn between trace metals .nd polycyclic acronutic hydrocaibons (PAlls) has attrneted increased irresesl recently since some Ir.ce metah, inchding seknium, have been shown to decrease chemical carcirror nesis. Due to the high abaorption e(ficiency of trace rnetals iw the hsng alveo)us.nd Me ability, of (wlmoaary alveolar maaopha6es (PAMs) to rn.iaboline PAHs, attenlion is certtering tww on the role of PAMs as a pnmary defense aRaiant teaobiotics. Foc the studies repxtod here, focus waa on the effects of aekwiww (r Na SeOJ and sulfur (as Na,SOJ on cytotoaic- ity a.d coejugation of bento(s)pyrex (B.P) by Aiwar PAMs. Rta.ks showed 11W, .Nfwrsh se k.iwn was fotind to be wwre cyroloak thr wlfrr, lhe cytotoaicity of both selenium and wlfur was greater for PAMa obtained by bronchopulmon.ry, lavage from wowsnwken than (nr thoae obtained lraw cipnttle amoken. At IpM sekniwn, ghr- tMMone conjr`atwrn of BaP was ewh.wced M bod wwoker and nowsmokcr PAMs Even though aeknwm roaicity cr be achieved itt PAMs at high levels (100µN'), it seems d+r this element may afford protection by i.dretion of detoaification enzymes. In- decd, die lack of sekniarn cytotoaicity, is swwker PAMs suggests a protective effect fram crganrne smote coeqo.ents, ind.dift the tr.ce metals studied hen:. Marsh.ll, M. V., Mcltwrxe, T. L., Srsb.r. O. L., Marrin, R. R., and Grif6n, A. C. In. Pofynrrfe.r A.owwic Hydoc.rbows F~G1 /nsen.arlowof Syw~otirrw cn Cherwital Andysis awI sioloticd Frt, Columbus. Ohio: Banelk Press. 1981. pp. 221 •230. OtAer s.qp.rt: The Robert A. Wekh Fardtlion and die American Can:er Society. From the Univcrsrry of Te aas System Cancer Center aW Baylor Cdkge of Medicine. I/ottston; Nonh Tes.. State Unirenity, Dento.. ONTt>CFNET1C VARIATION IN RAT UVER. LUNG AND KIDNEY Mt1r1(IOXYGENASE INDl1CT10N BY LOW DOSES OF BENZO(a)PYRENE AND CIGARETIE-SMOKE CONDENSATE 1n previorn in vlnn qtrdits. it has beat sbwm that aryl hydtocarbon hydroaylase (AHII) is specifically induced in the hrg and kidney of .ninul subjects by the inhala tio. of cigrette smoke, whertas ciprette snwke tus .o ttclioe oaAIIH atlivily ia the liver. These obaervNions were attributed to the roule followed by die cigarette smoke components after inlulatiow. However. the p.per prescntcd here, which deals with two sowes of administration. inhalation and i.p. injection..dds anothn parameter to the investigative pnuess. Whee administered i p., ciRarelle smoke cadcnsale (CSC) and benm(o)pyrene ( BP) in low doses reached the liver before the other organs. Neverdrc- less, thc long was still the most sensitive organ of the three. Resn)1s showed thN, in the liver.nd kidney. basal AHH activity (which is low in the (eqn)'acreaxs rapidly dter birth to reach the adult level two nqnlhs INer, and is only inducibk by CSC and low di»es o/ BP iw unweancd rats In the hrng, however. the basal Allll activity (ow in the (crw) urcrease. .lwlrly al birth. peaks in five-ay old rats and then decreases .IbRMly t'owrti w ewryme acu.uy in other tirsues, lung AHII canrrM be induced in un..cawed y.rnR .nrmal. tAc enryme subuVueMly bccumea sensdrve rn rndurmg a6enu and is IuRhly uwlot sPk in 90 day oW rats Srmrlar behavior has been shown to .W , occur in two other enzymes linked to cylochrome P,ISO: etlwaycoum.rie deedrylase and ethoayresorofin deethylase. According to the wdtors of this paper, the special AH11 inducibility of the lung of the aduM animal is. very important biological fact, and forthcomin6 studies should give insight into the cause and biological consequences of this phenomenon. Van Cantfort, l. and Gielew. l.E. , s.itisA 1o.vrwl aJCance. 4d:9o2-910, 1911. • OAs sapy..t: Fonds de Ia Recherche Scientifique Medicale. Frauw the 1Aboratoire de Chimie Mbdicak. 4witrl de PtlholoRie. Uaversit! de Lifte, LilRe, Belgium. ARYL HYDROCARBON HYDROXYLASE ACT1VfTY IN PULMONARY MACROPHAGES AND BLOOD LYMPIK)C.'Y7ES In the strdy reportod k,ae, the sinwN.wews anNysis of (1) ssyl hydrocrbo. hydroaylre (AIIH) iedretion is ctiltrned lymphecyln and (2) in s/rr levels of AHH activity is pulmonary dveolar nucropbages (PAMs) wr wrkrtake.. The study Rroap included eight Iu.R cancer patieMs with nbeMOs eapoare, 15 .o.<aaioer palic.r wNA asbes/oo espvsrre, 21 priew with lung cancer bM without asbestos tapoawe. ..d 40 patients with .eitDer hrwR c..cer .or sbeaoa eaposwe. All patients were ciRrene snakers who were .d,aiued kr i.dicMio.. of ptrao..ry disease. Wlw .N pnient grorps were compared in terms of their Individual capacities for tiawes to be i.ducad, s•rikinR differences were seen betweee tAe Rrotrps. Those individuaY with r+tEeatos eapowre presented increased AHII activity when compyed to those witho.t asbestos eaposYre whera lung cancer and aon-ea.cer Rroups were compared. Amarg cigarette senken with neither lun= eaucer anr oceetp.Honal asbestoa eaposwe, 6ft h.d telrively low AHH levels in both lyrnplacytes..d PAMs. When AHH values were eaasei.ed is asbeuo.<apaaed snaken widroul ks.R cancer, 57% of the i.dividr- ais eaMibiled k4w AHH levels iw bolh tiasres. When a uewllaeeoos cpep.rison wr t..de for cigarette smoken with lung cancer but withotr asbesws eaposwe, only 7691 o( dr individuals could be clasi6ed as having low AHH vN.es in both lymphocytes and PAMs. FiwWly, when AHH values were compared hr tissues oMair-Ad flom aabeskntaposed hw4c.noa pnieaMS the dMa showed tlut 100'><of these udiviArda poasesstd hiRh AIIH activity i. eidrer lymphocytes ar i. PAMs, bw wa is botb tiswes. Overall, these results suggest thr sdbeswa exposure re.y ef fed changes in a person's AIIN responsivenessor tolal induced kvel is sr.chew.y a a innease tIW iwdiv'd~iul• iul•s s wsceptibility to the materials fornd in cigreue smoke couder.swe. Swodgras, D. R.. Mclamore. T. L., Teague, R. B., Wray, N. P., aed Arsber, O. L. CHEST lOS:12S-/IS, 11181. , OtAri srqpsrt: American Caecer Society. Veterans Admiaistralion Hospital, Hous- bn, rd the National Instmrtes of IkaMh. From the Departmenl of BinkKicd Sciences, Notth Teass S1.u Univenity, Den1o.; tlepntmeM of Medicine, Baylor College of Mtdicine, and the Vdenms Adminirrs- Iio. llospitd, Houswn. I 8 9

M111.TIPI.ICITY ()F CYT(x11ROME P4S01N PRIMARY FETAL IIL'PAl (X.'Y7FS IN CULI URE The pmnnaW period of life is a critical time for the quantitative and qualitative development of marosomal monoosygenases, and earlier observations suggest that fetal hrpaoeytes in cuhure might constitute an ideal lool for studying the perinalal re6ulaatry mechanism o( mowoosytenases. Whik it has been known fa a whrk Ihat Primary fetal ral hepatocytes in c.Nve display difkrenl monoosy`enase activities which can be induced by sevenl chemical indreen. these hep.tocytes were believed wmi now to produce only one single eytodnome P-IS0 species, namely the cyto- chsorne P,450 (ar P4IS). Nowever, i1 now sxms possible b induce othercytachrome P450 species in these hePMocyses. prov1An they receive an appropriate hormo- .al teermenl. in the .rork tepated heae. es.rwinaliar was made of the effect of dearwcMaaone on various mowoosyUenres and on die type of cytochrorne P-I30 wrpporting these enzymk activities. ilree enzymes. aryl hydrocarbon bydrvaylase. ethosycounsrin doethy/ase and aieria suorwoayjalse, were measured (or this pur- po.e. Resrha of this study show IAaI die presewce of des.methnone in the culture Rr.dwm prodrees qudiW ive .nd qswiutive c#.n6es in the Inonooaygenase-srPport- is6 cyrocMane(s) P450. Fa krw desamedwo.e eoncentrstiows. acylochrome P450 is farnmd displaying biochemical yd biophysicr propcnies similar lo those induced by Phewob.rbilal in die adult rat lirer. At higher concenlstlion., similar qualitative changes are observod; but a quansiwive phesomenon occws. the (cylochrwec P4 50)- dependent enzymre s+ctivilics being also induced. Desametlusoae also has a syner`is- 1ic ef(oct in tLe indretion of enzymic activity by the miature of phenobabilal plus benzrafracewe. The various biochemical changes induced by desamethasone in die fetal cell cultures parallel those observed in vivo during the perinatal period of life. 7Aesefore, this eell culture system may cawrlule an interesting model (or studying the ootorsyc development of liver monooaygenases. Kremers. P., Coujo.. F, De Grseve. 1. Van CrwfoA. 1 and GKf.n, /. F. Eavapean lowwal oJ eiorhtrnutry 116 67-72, 1411. OrAar sawart: Fonds de la Rocherche Scieatifique Mldicak. From the Ls6onloire de Chimie Mbdicak, lnstitw de P.tholol~ie. UnivcrsNf de Litse. Lilge. Bdgirwu. DNA METNYLATION IN NORMAL AND SVIO-TRANSFORME() }IUMAN FIBROBLASTS The 5-methylcytosine base eontent of DNA in fow norseal and fow SVIO-trans- forsned hwn.n diploid 6brobldt twkrres was measured by high performance liquid eMorn.a+jraPhy (HPLC) ResuNs show dr1 the percenl o(cytosines methylaled for the (uw nnnnal cell lines ranged from 2.23 to 3.16, whik the range far dre (nur SVIO- transfarrrrd cclt% was 6nrn 2 90 lo 3 03. The mean frx the total number u( IIPLC drrrrmrmuuns w as 2'14 • ll 29451 dclerminalMw+s) frw the mxmal cell types and 3.00 t 11 211 /11 dcrnmmarNrns) lur the Irsns(ormed hnes Thus. in cuntrasl lu other reported studies camparin6 normal and oncoseAically transformed cells, no spp.renl difference was observed in the 3-melhykylosine to cytosine base ratios in the two ceU types. 11 is worth emphasizing that dre IIPLC method used here gives an absolute measure of die DNA bases. In addition, the plrrNy of the DNA is controlled by monitoring for the presence of wacil. Other methods using radioactive label may be hindaod by. various differenl artifacts. Diala, E. S., Pkat, M. M., Coalsan, D. W., and Hoairwn. R. M. Bioc/lrrnicaf awJ eioPAyskal Research Cowrnrnkorions 102(1):U79-13LN, 19%1. OrAer salip..t: National Institutes of Health, The United Cancer Cowscil, Inc.. The Cancer Researcb Coordinating Committee of the University of Cdifarnia, the Aca- demic Senre. Univenily of Cdr(orni.. Sam Die6o, and the Leukemia Society of America. From dse Department of Podiatrics. Univessity of Criforwia at S.n DieRo School of Medicine. La loll. I CONS7TTl171VE BEHAVIOR OF METHIONYL-1RNA SYNTHETASE COMPARED TO REPRESSIBLE BEHAVIOR OF METHIONINE ADFNOSYLTRANSFERASE IN MAMMAUAN CELLS Mcdrionine, becwse of its roles In pralein synthesis and in nsethylrio., b of tentr.l inpartas,ce to a!1 cells. In the metabolic proeeu, aredlioeine can be wed by the oeU Ilrwth two different pathways: (1) methioalle can be eonvesseA so S-adeooo- sybselhionieen die Inyor mnhyl source for eepdsr twmethylatiow reactiown and 1he w.ce of the propyl.mine group for polyanwne biosyndrcsis. or (2) it can be convened to meMriowyl-tRNA, an iaportrM iaesntediae in p.olein biosynlhesis. Is the paper p/eseMed here. il is reported thM anethioeyl-1RNA sysWretase, unlike loethioni.e adenosyhrwferase. beh.ves in a cantilwive mrlner with respect to the concentrsrion of inethioaine iu the culture anedirla. lllis behavior Is see. in Chinese h.nater ovary cells rd in normal diploid and SVIO-trmsforsned brre.n fiboMasss. Although die kin4ics of regulation of arcdliowine adewosyNrawsfer.se and anethionyl-IRNA synlhe- tase by esoUenous mclhiowine are cleady differenl. the levels of the two enzymes hl lhe hwn.n cell lines re simibtt. Rrbnilz, l. E., lacobren. S. 1. and HoA4wan. R. U. Siochiwlca er eioDOiyska Ac1a 677:2d9-273. 19111. OIAai aarr..f: National Instilules of Ne.Nh, The United Cancer Council, Inc.. llro Cancer Reseuch Coordinating CalMnitlce of die University of Califonlia, the Aca- demic Seaue. University of California. San DieRo, and the Lerkemi. Society of America. Prom die fk(wtment of Pediatrics. Universqy of California at San Diego School of Modicine. La Jolla. ' 13 12

F!)I.AII? P(H.Y(iLt1TAMATE AND MON(Xi1.UTAMATE ACCt1Ml1[ ATION IN N()tMAI. ANI) SVIO-TRANSFORMED /II/MAN tIBR()BI.ASTS In the peseM aaempt to ascenain the role of folate pdy6lwamates in cell divi- swrn, it seemed neccssary hrst to surve the cells of (olates and estimate their irMallydNe requirements fnr growth. All fnur ceil lines studied here, the mxmal human drpluid fnreskrn hhn"ast PA, the normal human fetal drPloid AF2, and the SY40-trans- (nrn+ed bncs PS and P1, showed sinwlar kinetics for falate starvatiar. SepFadcs C-10 gel hhratwws chromatography was wed 1o wrcawre the accumulation of fdaoe prlyglu- tamate and nwwwoglwamate in all of these cell lines. After the celis had been depieted of fiAates, they were provided with IinMins aero.nls of (711-fulic acid in order that the cells would accumulate only forms of folale nxessary for proliferation. Bah Ihe a.rnal and the transformed cells accrnwlaled monoglwamate and prly6lwamate forms, hw by 72 hwrs of labehn`, the transformed cells cvuMained 3 K) times nwne p>ty~lwsnwe then the nmmal cells The Rrowlh raes for the rwwmd and IanslrMmed cells were similar at this limiting folic acid eoncewatwrn. Thus, rf k>1Me p4ygluta- nwes we mnee rmpwtw for the prolrkrsiinn of SV141ransformcd cells than the wursnal cells,lhe inMbitiow of polyglwarwre fornsriow could possibly be an impextaM potential trger for chenwuherapy. Ho,/fiwaw- R M to ol lewwol nJCrllolar PAysioloRy 109:497-503, 1951. OUAn sayprr: Narwsrrd Instnwn of He.hh, The United Cancer Council, Inc., The Cancer Research ('oordrnatrng Commmee of the Unlversily of Cahforria, the Aca- demic Senate, University of Cdrfornia, Sam Diego, and Ihe Leukemia Society of Amerrca. ~ From the DepartmeM of PedrMrics. Un/venily of California N San Diego School of Medicrne. La loll., and the (ienetres Unit, CMldren's Servrce, Massachusetts (kneral Ihnpital, Departmenl of Pediatrics and Center for Human Genetics. Harvard Medical School. Boswn. DNA METHYLATION IEVEtS IN NORMAL AND CHEMICALLY 1RANSFORMFA MOUSE 3T3 CELLS TMs {nvestiRation was undertaken 1o assess the effect of chemical transf.xnution on tntal genomic DNA methylatwn as measured by high p:riormance liyuid chnwna- aWq+hy IIIPI.C). In the study presenled here• norsoal mnuse embryo l7l cell cultures nrwl dwse oncoRenw:ally transformed by the chemical carcinosens benrola Ipyrene and methykholanthrene were andyred by HPt.C 1n determine the S-me11 ycytosine to cytosine base ratw» in their total genomic DNA. Results showed that thc mean fur 10 11 W.C detersniraliaa of the normal n) cells was 2.87% of eytosines melhylated with a standa.d deviation of s0.Se, while thM for the benzola)pyrene-transformeJ 3T3 cr lls and the mnhykMrlaMhtene transformed 3T3 cells was 2 99% ± 0 N(11 dclermi- n.u..n.l rw1 2 NI's •// IN IIS ikrermreatwwxl, resptctrvely llrcu reudts led to the i.ww lu.i..n rhr itwic rt rw- rrd ddlcrcnce rn the e.rrnt of wwal Ecmr.na• I:NA mrthyla- r..o t.F..«n mrnul .rd. hrmw all) uan.lown.rd 11 1 icll.,rhen mc..wcd hy 1/1M C I Diala, E. S. and llroffnrsn, R. M. ' Aioclkrnr,nl mwreiuqrAysicnf Resrur.AConunwricwiuns 104(I):11d9 1491, 1982. OrMer rrhert: Natinna11ns1itules of Heahh, The United Cancer Council. Inc., The Cancer Re-earch ('.rsrdinatin6 Conunetee of the Univenity of ('altfomia, 1he Aca- demic Senate, University of Cdifarna, San Die6o, and the Leukemia Society of America. Fnwn the Department of Pediatrics. Univer%ity of California M San Diego School of Medirine, 1a lr+lla. MF.TIIIONINE [W.Pl:NI*-NCE IN CANCER CEI1S-A REVIEW Methirmine dep:ndence occurs in a large rwmber and wide variety of cancer cdb a.d dnes arw seem to be a r.ndorn eomponenl of the 4tnsforwted phenotype. De6nitio. of methionine dependence slales thM it is a defect found in many cancer eell liwa 1hM inhiMts their growth in culture when methinaine is replaced by its immediate prec.r- rx, hurnorysteine, in the culture medium. Norsaal c'uAwed cells do no1 have this defect. This report lists the diverse and large nurnber of animd and human cancer lines thr we methionine-dependent, and critically reviews the cell brotoRy ud meMioeine bioehemisuy of the plKwoarenon. Haf/iwaa. R. M. In VMre 1i(3):121-12f, 19R2. Otlrer sappert: National lnstitwes of Health. The l/nNed Cancer Counell• Inc., Tlr Cancer Research Coordi.riwR Committee of 1Ae University of C.fi(orui., Ihe Aca- demic Senate. University of California. Sr Diego. and the Leukemia Society of America. From the Department of Pediatrics. University of California N San Diego School of Medicine. La lolla. HYP(NMETItY1.ATION OF IIFLA CEI.t. DNA ANl) THE ABSENCE OF S MI:1'11Y1.CYTOSINE IN SVIO AND ADENOVIRUS (TYPE 2) DNA: ANALYSIS BY IIPLC in the study presented here, methylation of 1he prihed virion DNA of bnth SVIO and adenovirvs /typ: 2/ was mcarired by high perforsnance liquid chromatography (IIPir) and compared atheir Mrsts, African green monkey kidney cells and HcLa ceNs, respectively. In SVIO DNA, as much as 12 nanomoles of cytosiee has eeen measured without cwrcomitad delcayion of mCyt. SVMI contains 27 Cp(3 pain. which is the usual methylatam site. If all Cp( j pin were methyla/ed, this would yiekd a siRnrficant 1. )'16 methylatiun of L'tal cyhnines; however, in the virion 1)NA studiod here, nmwa scemed to he methylaed Also, as with SVN), mY'yt was nnt peseM in the IS It

.1 1 IN the com-lusion that candage contains estraclabk matrss compounds that inhibit inva- sM"n tn an eapcnmeMal system When these dilfusahk and estractahk sulnt.nces were lunhcr studied. it was found that the rnhibtian of osausarc.wna cell prt IderalirMt was caused by molecules with a molecular weight of less than 50.4100 daAtins 1KGsm this anu invasive facair of the cartilage estract, a pruatease inhihitur was tdaNilKd thal has the aMtbty to inhibit mamalun collagenase. including that elalxxaled by ostunarcoma cells and endothclial cells These esperimental data led to the hy(w+thesis that invasion cof cither tunrir nr endcwhrlul cells depetds on praeulytic /a+llagemdyticl enzyme n tivNrcs Other studies aktns these lines have been instituted to follow the invasive- ncss and prohferation of bladder eancer. Kurnrwr. K. E. and Pauli, B. U. In Gdbcrt, H A, Weiss, l.. and Monsew, D C. G. (eds. )' Aowr Mctouotn, Boston: (: K Hall Medical Publishen, 19l1, pp. 1)1-163. Other sayla.rt: National lnswitutes of Ikahh. From the Departments n( Orshopedie Surgery. Paitholosy and Biochemistry. Rush- Presbyterian-St. Lute's Medical Ceneer. Chicago. ANTIINVASION FACTOR MEDIATES AVASCULARITY OF HYALINE CARTILAGE To test an arMiinvasion factor IAIF) hypothesis of the resistance of cartilar to vasculr invasion, a novel in vitro system was studied Iha1 employed bovine .rttcular carttla6e aa a growth surface (ot normal heparin-ssimulaWed eaddhelial celis. In this study, cells were tested for their ability to invade the malns of viabk and devitalized earacted cartilage as mrwutortd by thin-seclion electron microscopy. The growth behavior of cells on devitalized estrsetod cartilage was esnnined in the presence ard abaence of cartilate-derivedc estrsclable AIF in the culture medium. Whereas wur- ' mally viable att/cular cartilage is a poor growth surface for Cndolhehal cells, the eadnthclial cells studiod here. is eataasl. grew as conlact-itJtibited anomslayers of Ilaaenod cetls on the surfaces o( estracled cartilage. The cells were separated frorn the carulage tnauis by abwtdant bwl lami.a. There were a few micnoviUi at the basal ptasma teembra.e, but there was no de:Sradatioa or penetration of the eolla6enous snnris of eatrat:lod cartilage. Hosvever, wAa e.dashdid cells were stimulated by hepari., they assumed a polyheAnl sh.pe and Pc^tlr>NeJ the estracscd cartilage with awnncroas microvilli and some eyloplasmic processes. 'tlws penetration of Me colla- <enous matru was associated with tlswe rarefaction a.d degradation of eollagett fibea. heportamly, however,llus invasion of hepari.-stimulated endcMhelial cells was .bdished when (ow conceatr.tiom o(e.nilaje-dcrived AIF were addcd to the culture medium. These data provide evidence thN the resistance of hyalame cartilage to ee- dadhelial ccll invasion is resulaled i. patt by tissue derived pnNcinase inhibipxs and a. sntipmlikrMive activity directed atainst endolhelial cells. Ku.untr. K E. rt o/ Seiwinors in ArtMftis A RAera.otiun 11:67-69, 1951. Other sr.*pr1: National Institutes of Hedth. fnwn Rush Medical College. Chicago. [E I C1IARACTFRIZaT10N OF ADULT BOVINE ARTICULAR CHONDROCYiES IN CULTURE AAiculr cartilage slices, obtained (rom IR-naold bovine me/acarpnphda.beal joi.ts, were used as sowce mNerial for thia descriptive article of chatdrocyla in culture. After sequential digesliowt s.d 6hrMioe, cells were plated in either tissue culture dishes or roller bottles. Chondrocytes f ned iu buffered glulraldehyde corai.- ittR 0.1S svthenium sed, were esamined by Ii`M and transmission electron micro scopy. Collagen type deknrtin.tipt of 'H-proline-labekd proteins isolated from cultures were performed by elecyrophonyic atd CNBr peptidt: analysis. Biosymhesis of praeoillycans was measured by »SO, iwcarporatiom inso wwromolecuks eslractod under dissocialive conditions. Eaatei.aion sMwed qu1 isolakd clwndrocyles prior b culture were typically r.otrried with scaM tetriwrid mrris, which could be Rmoved by mild trypswrzation. TMatghout IMe prognession o( drc euNures, phenotypic .heq- 1iow. wete .ot obser.rod. E(pclro/luorop.plp of eo11" thal was entracsed from'N- pmline-labekd cuawes aflet nwW PeVundigesoionshowed i band i. drc positio. of the al chain; an e2 ch.in caad rwl be delccted. Cya.oRem bbaeide peptide analysis oblaiaiod 6wn type 1 mlor radioacyive peptides eo.wirabd with unlabeled pcptides ype collqe.;lype I eollagert was taot delecublc is these cultures. PsuseoglytM aggregate was eatracted fran both culture disAes aad mlkr bottle cul- lures, under associrive conditions. Thpe wem diffese.ces eoted in this esperannr suggesting that tfrc roller e.Uure eca-associaled ananis may have a pener depee o( organization tlun that pows in su.0ard tissue culture dishes. These same data also indicate that anicular dqndtocyles Rrows is mas ro(kr cuhures we capable of syathe- sizi.6 a phcnotypically stabk, tissue-like nulri: in vitro. Xartawr, K. E. ed at. Se.wawrs iw ArtArit/s t RAnreoHaw 11:101-10), 19d1. Other npport: Nriow.llawitutes of Heddt. From Rush Medicd College and tlte U.ivenily of Illinois Deet.l Schoo(, Chicago. REGUTATIUN OF TUMOR INVASION BY CARTIUIGE-DERIVED ANTI-INVASION FACTOR IN VITRO Manunaliam crtilaV is highly resisunt to invasion by tumor cells. This asiss- ssroe was swdied here wi/h 1be use o( a oovet Jn vlno culswe system. Aniculr catilade obuined rsan fresh metacarpophdanteal joints o( preadolesceM bovines was used n a growth surface for huma. TE-IIS osuosaRonu celh and foreskin flbrohlass. Cartilage d'ats formed the boltoms of sui.kss-wteel cylinden, providing closed growth chanm- ben Gor these cells. Both invasive osKOSUroma cells and normal FlrroMasrs were tr.abk to pennr.ne viabk, uneatracad catilage durinR a 2-weet cuNwe peaiod Whe. canibge was devitalized by freezing and duwiqg, die tissue remained resistant b invasion. Cartila6e, eslracled with either I or 3 M tuanidine hydrochloride, was invaded by oueoaarcoma cells, but mA by control fibroblasts. Invasion by osleo.r- eana cells into saN eatrae/ed eartila6e was abolishod whea low concentrations of a cartila`ederived, atqi-invaswon factor were added to the culture medium. These dNa 19 ,r

_L provided evidence Ihallhe resistance of cartila6e to Iumrx invasion is regulated in part by tissue-rkrived pr.wemase inhibAon. Pauli, B U., Memoh, V A and Kuetrnrr. K. E. hwrnof oJ1Ae Nurr.NSOI Cone er lestitute 67(I):6S•77, 1991. (kAer r.yw.t: National Cancer Institure. FnNn the f)cpar(ments of Pathnlogy. Biochemistry and Orthopedic Sursery, Rush Medical College. Rush-Presbyleria.-SI. Luke'a Medical Center, ChicaSr,. IN VfTR() DETERMINATION ()F TUMOR INVASIVENESS USING f:X1RA('TI:D I/YAt.INf: CARTILAGE In this anenspl to determine whether sah<atracted cartilage could be u sed as a tesr connective liuue for in vitro discriminaion between noninvasive and invuive tumor cell hna, a novel in vitro nsethod wY devised which used salt<suacled, bovine nucular carlilase as aa esperimeaAal Rrvwds surface /or both normal bladder epithelial cells and rwiwinvasive, invasrve, and mefastalic carcinoma cell lines derived fnrm chemical carcinogen induced tumors of dse ra1 urin.ry bladder. As monitored by Ihin- aection electron microscopy, sah<atracsod cartilage was readdy penetrated by the invasive and nrUstatic rat Madder carcinoma eell lines. The metasrurc cell line could be differentiated from the invasrve, noametauric cell line by its grea.et depth of InvasNM In ccrwrast, noninvasive carcinoma cells as well as normal blrkkr epslhehal cells lacked the capacity to crrtsde and penetnte the eatracled mtlris of the articulr cartilage IIsrn6lhesedefinedcell lines,sah-ealrnctedcartda6ecanbeusedtorepaxlu- cibly discriminate between can uamas having different invasive polentials. This assay system may have diagnostic application for the tn vitro staging of lumura. Pauli, B. U., Memoli, V. A. and Kr.rtnwr, K. E. Cancer Research 11:2061•2091, 1981. OOther sayl.rt: National Institutes of HeahA. Froru the Departments of PaUsobffly. Biochemistry, and Orthopedic Sur6ery, Rush Medical College and Rush College of Health Sciences, Rush-Presbyterian-St. Luke's Medical Cenrr, Chicago. A FAMILIAL AGGREGATION OF PANCREATIC CANCER: AN IN VITRO S7vDY Pancreatic cancer, with its olncwe etiology and diffscuh early diagnosis, presents a medkal problem of slaggenng propurtions. In the in vitro study presented here, a family was identshed in which four individuals manifested pancreatic cancer verilied through two generations Cell cultures from split-thickness skin biopsy specimens were nblauied frr 24 rnembers fnsm three jenetNions (17 bMrdlirse relatives, seven family memBen by inarruge) as well as len noefamily normal subjects, none with a lamily hwory of aild sunrxs One of the conslam features of human nwNwolayer dcrmal (ultunrs has been dipN»dy On the other hand. hypenhploNly. other than 1et..pkidy. has been rarely observed in cultures fmrn.ormal subjccts without a family history of solid tunwus. In this shdy, however, increased in vrtru hyperdiploidy was observed in eight of the 17 family members /esled. In the long run. allregates of pancreatic cancer in families such as this one. and the occurrence of pancreatic cancer in sonse .wosornal domiarst cancer syndromes have added credence to the relevancy of agenetic component(s) in a fnctioa of patscreMic eancers. ln an important way, this study has demonstrated the potential lnr eombirdng detailed family data with rocogni- lion of in vitro biornarters for caucer pronencss an am approach ao rhe cornpehension ol carcinopeesis in pancreatic caecer. D.nes, B. S. and LyncA. H. T. IAMA 217(20):2798-21102, 1982. Od.r nff.rs: Naliond lawiluks of Heahfi, Danes Medical Reaeandt Fu.d, Cor.cll lhdveairy Medical Cdkge, and ?.emwray Foundation. Fruas Ihe Laboratory for Ce11 Biology, Department of MedKine, Cornell University College. New York, and the Department of Pmventive Modicine/Public IleaMh, Creighlo. University School of Medicine, Omaha. GENETIC/EPIDEJNIOIAGICAL FINDINGS IN A STUDY OF SM(NCING•ASSOCIATED TUMORS In dsis Rendiclepiderniobgical sludy family histories of cancer were obuinod via personal interviews frvre eonsecMively.scertained eancer patients who were under evaluation in one of two University Oncology Cli.ics in Nebraska. Inchrded is the srriea were 147 breast canetr pobands, 85 eolor cancer proe.nds, U lung c..ca probards. and 111 prahnds with other casmen Yot have been n:patcd b Ee asocired with cisarelre smoking (carci.orna of tlse oral cavily, esophaRtts, p.aereas and uriaarp bladder). Smoking histories of pwbands and their relarives were obtained for an oveslappinR series of 60 hrtg ea.ces prob.nds and 7111 psnbands with other smokiy- wociarod romors. Findi.p revealed tlw, aNho.lllt a significam cohort effect was observed with respect to smoking habils for bah relalives of lung cancw probands.nd for relarives of probanda wilh other smokinR-associMed rwnors, a corresponding arewA far hsnt cancer frequeney was observed only for relatives of lung cancer probar,ds. This resuN suggests the importance of host faclon in consbinalion wNh environme.ul eaposures in determining hsng risk. A cohun treed for hrsg cancer was also appareM amons reftlives of breast cancer prolwsds, but not for relatives of colon caarer pro• bands, suggesting the posublNy of an intrlnsic association belween carcinoms of the baast and lung. It seems reasonable that further ehrcidaliun of hosllacqor suseepribil- iry in hmg cancer nsay have importantetiolu6ical and preventive implications. LyncA. H. T. et at. Cancer Genetics and Cytosenrtirt 6:16)• 169, 1952. FrMrs the Inslilule lor Familial ('ancer Manasemenl and Control, Inc.. Departments of Prevenrive MedicinetPuMic Ikalth, Surgery, and Pathology. CreigMon University Schkal of Medicine. (Mssaha. 20 1 21