Tobacco Documents Online

HT®0120107 C In Vitro Assays The pmedin$ section showed that tobacco smoke contains chemicals which interact with the mpnooxygenase system. This interaction can be beneficial if it produces nonbiologically active. polar metabolites which can be excreted from the body, or it can be detrimental if it results in inter- mediates with exceptional biological potency. Table 2 shows that IAI CSC fFactions are metabolized by enzyme fractions from rat liver to fortts mutagettic to tester strains TA1538 (25. 27) of Salnu,nedla ryphimuriurrt. and also by endogenous cellular en- zymes to forms capable of transforming the C3H IQiyA cells in culture. This table also depicts the levels of nicotine, phenols. and BP in fractions Bf. Wk. and Nv„--the Bactions which contain virtu- Tffi1s 2. F.flkm of 1Factlem of 1A1 d0.alena amWiO caade®ee ICS>:) tn .aibus meAd sI .. n. 'xkBP •a ~ OItVoOIlM 0o/MN/N, Oa1010 OMt070. . ! SA„. actually inhibiting AHH levels in the lungs. The fractions that induced AHH activity Ie.g. B,a or B,bl contained chemicals that could metabolically compete with BP in an in virrn assay (see Table 2). This implies that CSC contains not only chcmicals that induce pulmonary AHH. but also chemicals that are potential subserates for these enzymes. What Are Some in Vitro and in Vivo Effects of TobaccQ=P!®I®ted Chemicals? :.~. . . . . . . . . ., .: .P... ... w ~ i 0 0.1 05 t0 wa, 4CnvnY ,unrtsro nBSUL1011111 Ftoune :. Potmosary mrtpoase of B61)2F, x D2 proeeny to nodt MC end (wnoml waste ctOerom smoke. Amatds were 8erw erated and houttad as dasenbed ttefote U2). terocul®uan of MC and smoYs exposure was given at daenbd in legend to FtBUre 1. AHH sstivay was detennltesd 24 hr after R Instills- Han of MC and 6 hr after expowte to ci0etem smte. Mtun and stendatd devlmdon ts gi.en for tho MC•uestetl popule- tinns tAif am teproaenmrive of tNe nomesponsive i0.23 e 0. 10) and rost>onsive (0.95 e 0301 poWlations. nomenclature were as described by Swain et sl. tl6). Table 2 summarizes some of these effects. In- trauacheal admittistretion of the whole CSC and re- constituted fractions induced a stnall, but sig=tifi- cant. increase in pulmonary AHH activity. The CSC fraetions varied in their capacity to induce AHS activity with the frardons B,a. B,b. Nv,,,n. and N,,, the best inducers. fractions Be and WA, wealter inducers. and fractions B,,. SAt. SAr and Frssnod Content. ntBag AHH inducnbility' to pw SOab mhibitiort Mulagsnesitt Trass(oremtbR Wlwk CSC 23.30 1.7 3.0 +-. . tteeonwnued CSC 23.00 1.8 !.2 B,n 0.81 3.6 0.8 B,b 0.29 2.5 0.5 B, 0.93 1.5 3.0 B. 0.36 0.3 >10.0 WA. 2.27 1.6 5.0 +• + WA, 1.98 1.1 2.0 _ - SAj 0.39 0.5 s19A SAr 0.78 0.3 >10.0 - - SA. 8.69 0.4 >10.0 - - Nr„s„ 1.19 2.5 3.0 s - Nrr 4.38 1.2 ND - - N,,, 0.70 3: 1.0 _ - ' Whole CSC has 21.0 mB nitotinarg. 5.70 in8 tibsooler8. aoA 0.98 p8 BP/g. Ieeonsututed CSC hae 22.0 me aicodem8. 33 m8 ptawtd8. and 0.90,.r, BPrg. Bn has 31.0 nm8 moonna8. WAir hen al n+8 plronolerg. and N,,,, hu 31.1,s8 BH'a. Datai4 81ven by ttatet ot d.12!). ' AW! indacibdity - aBeet or Itnations of IA I CSC ttsetbns aa pahaomry Alifi ectivtty of CS7BL6 Cmn mfea ralsWre to a atqe n8 aam01 uI). ' aP inAibitfoe - wmpentlve tn wr.o atTect of CBL Oe:rnans of BP metebolLm by hopstto mocrowmes from 3-metbtrkholumbrow tleeted CS7BL6 nrda .U31. • MuMPONs s muttaaaic eaivny of IAI CSC ftaWns in SoheoN.Ba typlumunnnt T.t 1S18 m the gmastxe of Ihrer miaotonul S-9nWtti01. ' TtaoslcnneHan - malignant unosfonnonan freqnrecy in CH 10T1C Cl. 8 calls treated with C5C koetpns OF). Apsil 1979 65

u=~r..,_ . _._._._ ..____.~._ . . _ _._.... _ ..-,o-..~.... HT00120108 ~ ally all of these three chemicals. The most bioloRi- cally active ftuctions are not those that contain the BP or phenols. The Bra and Blb ftactions have not been analyzed for their chemical content. However. because of the nature of the &at;dotustion scheme IB,a a bases insoluble after ether. B,b - buses in• soluble before ether), these fractioss could contain some aromatic amine-lite chemicals. A similar distribution of mutapsic activity was found with CSC ftactions from the 2AI reference ciBarette (Table 3), and such activity was observed to be stable even if the fractions wrre stored frozen Mutmu par pletm Pu730µgot TaE6o 3. Msmp=alf oiTA 98 dID 2A1 elgartsti cmft amdmzto ltSll nosdwa.• Coeeem, Praetme movie ubefe CSC 40.00 Beaoafmuozd CSC 39.50 er 0.60 BP 0.24 BE 1.06 B. 0.32 WM 3.63 aA% 2.ti6 SA4 1.44 SAM 0.88 sAw 17.60 ~ N~ 2.16 ~+r 9.34 Nw 1.24 ISUMPIO for one year. Thus. for at least two different types of cigarettes. cer.aitt fractions of their condettsgtes can be metaboliztd to forms that are active in an in vitro lautagenesis test system. However, subsequent studies (Table 4) revealed a potentially important hindrance to the extrapoiation of the foregoing studies to an in vivo situation. Mouse Pulmontary tissue preparations, under con- ditions where BP is significantly sletabolized to 3-OH•BP (see AHH levels). failed to significantly aetivate either 2A1 coltdensste or t.aminochrysene 16AC1 to forms nsutagenic to the TA98 tester strain. Glculeim mumars WPMOm algflrerte Acpvuy.% [sxq I B+tpl 2 11911 8up 2 8ept I Eape 2 133 122 21,280 19.l20 179 148 2l.1=2 23.214 967 1.149 SJ69 3.758 11 10 3310 l.266 3.370 5.035 16 to 1,258 2.00S s.93 8.790 2s 30 102 Isl 212 314 1 1 2119 394 4.196 3.72.1 m 20 141 :05 1.lOg 2.192 7 8 53 43 30 248 2 1 8 14 28 49 0 0 8 20 499 1.248 4 65 93 M2 664 i 3 A 24 2.17s 916 10 3 Bo 139 397 689 2 2 • Thn 2A1 Uow aeeooml CSC 6scuom werc genatmed try Meloy Iabwemnes aceordia8 to ths rmtMftB of PWoI at e!. W1 a•A the Pow plate utcorPomNVe mm4gmem asray was petrmmetl accordhg to ihe mal4odi of Kier at aL (241. Tablo 4. Cspae{tP of TCDWndumd moum tqgstle sad oulmaaar/ S-9. a.d %nebr 1iSadndooed rat tw"Ma 8-9 to ntlvmtp w0ptp efgrae ®ote coodonne to fbtml.l m- w I I to S. qpAdwari.nr TA 9B.• Compound Soutoe of 3.9 TYPe +-WF, le CS7BL6 hepaua PM Pmme 4M. Sueenvnl. Musmwpmte Piotao. mg pmolatube" Gfd Immus taclienamdl 2A1 630 3.14 .939 IIS AP% 1 3.14 4919 170 2A1 260 3.14 4929 lo! 61 6-AC S 3.14 4929 102 11s Cl7BL6 polmvaary Pmr plete ,2A1 1300 11.5 8800 8 AFB, 3 11.5 S8011 Lte Suspensioe 2A1 1300 1.4110 1071 105 ! 6-AC 123 6.90 3208 83 3 Rat heperk Pow plate 2At 130g 4.43 9623 1g! AFB, 1 4.43 9627 760 soa9eoadon 2A1 63g 2.93 7371 106 83 6•AC 0.5 0.03 190 82 3» ' Pow PW toci'Pot mumg- asa®ys weie performod escordng to the metAod of Nesr at al. IIe1. suepemlan esmye aotmWad of n 35 mip ,ncuba[don of a muuue of beoorm. 5.9. and ua chsmcaU ia a tsutfer composed of 3.6 mM NADPR. 6.2 mQt NAAH. 3 mM MpC7,. 8.1 mM N4HPO« 13 mAf KltrPO„ 2.7 mM KCI and I37 mM NaCI. ' Ae p®ole 3-OH-BP fumed per way mbe: total ume wot 35 imn: 25 µg BPmI wau suUstraee. ' Peieemege of tuamm wwivulg 3! min suapenaron reladwe to TA 98 al,mQ. ' Subtequent esisYa with 11.5 mg prosem. 8800 aqiu AHH. and 1300 µg :Al stl8 ihowed no induced rnusanu. *I Enrfwetnental Heallh Perspectivrm I

, HT0,012011( ( I Addition of ferric oxide I FeyO, ) with BP resulted in 8°!c malignant tumors Li0). However, treatment with selected :AI condensate fractions every other week for the duration of the study 139 weeks), resulted in significantly increased malignant lung lesions in animals treated with both BP alone or with BP plus FerO,. The fractions alone. or with Fe=CI,, resulted in few malignant tumors 0 out of a total of 764 treated animals). Thus, in an in vivu lung tumor model system, the fractions seemed to have mainly "promotor-like" activity. Can Individuals or Tissues within an Individual Differ in Risk to 4ho Effects of Tobacco-Related Chemicals? The answers to this question are at best complex. The results of studies discussed in the previous sec- tions can be somewhat difficult to reconcile. On the one hand. tobacco smoke contains chemi:als that interact with hepatic mixed-function oxidases such as AHH. and this metabolism generates inter- mediates that are mutagenic and transforming in in rirn, test systems. On the other hand. CSC is not activated to mutagenic forms by mouse-derived pulmonary tissues in tvm,. Also. CSC material does not seem to efficiently cause lung tumors in rira. Many mtetpretations are feasible. Perhaps the lack of activation is characteristic of mouse pulmoaary tissue alone and cannot be generalized to other species. In point of fact, rat pulmonary tissue Fa: been reported to aStiv tte certain smoke :onden- sates to mutagenic forms (24). Huwever. both rat and human lung tissues have also failed to activate CSC material (24. ; l). and so the generality of the activation (or lack thereof) is still not understood. Second the fact that hepatic tissue can activate these condensates suggests that tobacco material could be activated by this organ and, following that. the tissues at risk (e.g.. lung) wculd be exposed to these active Crsnns. Or third. th~ lung is at risk only for the effects of those chemicals in cigarette smoke capable of "promoting" the carcinogenic event that may be initiated by a mynad of environmental pol- tut9nu. The first interpretation is difficult to assess be- cause no lung cancer model animal system directly appli;able to man is currently available. However. the kind of pulmonary lesions observed in mice fol- lowing exposure to known chemical carcinogens, that is, bronchogenic squamous cell carcinomns, al- veolat' adenocarcittomas. and adenosquamous car- cinomas. resemble lesions found in man (,f?. 33). Some data suggest that the liver plays a major role in the eventual susceptibility of other organs to chemically-induced cancers (34). However, no evi- dence is available to suggest that lung tissue is at risk from liver-metabolized chemicals. Evaluation of this alternative in t'iwr will be difficult because of the close liverrlung relationship. The third interpretation is panicuiariy intriguing and suggests that "promotion" represents the real risk of cigarette smoke to lung tissue. This idea is consistent with the following facts: (a) tobacco smoke condensate is capable of "promoting" mouse skin carcinogenesis: (b) tobacco-smoke con- densate can "promote" lung carcinogenesis in model animal systems (see Table 6): (c) the condi- tions of cigarette-smoke exposure that result in the highest risk of human lung cancer are quite similar to those that are most promotive in animal test sys- tems: that is. frequent and relatively prolonged treatment: (d) the chemicals in cigarette smoke (particulate phase) that are known inifiators of car- cinogenesis may be too low in concentration [total m 400 ngrcignrette 4>iS)1 to initiate sigaificant ttans- forntation in vivo. The evaluation of whole cigarette smoke as a potential initiator and promoter of lung cancer in the inbred strains of mice is now being studied in our laboratory. NOTE ADDED IN PROOF: We have recently folmd that a pulmonary S-9 preparation from Aroclor 1254-induced mice is capable of weakly activating 6-AC to a bacterial mutagen with the use of a pour plate assay 12-3-fold over background). We have still not observed an increase in bacterial mutations using this S-9 preparation and 2AI cigarette smoke con- densate. This study was supported in part through contnicn fiom The Council for Tobacco ttpeereh Inc.. U.S.A.. New Yort. N. Y. The awhors thank Ms. A. Venture and Me. C. Dwyer for their asurtanrx in typmg this mamucnOt. SBFBRENCBB 1. Lynch. H. T. Cancer Genetics. Charles C Thomtu- Spnr1 a Iu.. 1978. :. Mo+on. T. J. Atlas of Caneer MoedIty for U. S. Counties: 197Q19ti9. U. S. DHEw. Publk Health Servrce NIH. aethesda. ANd.. 1973. 3. Lynch. H. T.. Guirgis, }i. A.. and Albms. S. Famtlial ussor,- atron of carcmoma of tho breast and ovary. Stug. Gymcol. Obrrct. 138: 717 (1973 1. a. Knudson. A. G. The genetics of childhood cancer. Cancer 35: 1oL' i197S). f Petrakn. N. L., and King. N. C. Genetic mtukers and cancer epidemiology Cmcer 39• i8o1 ,1977). l 68 Environmental Iteaith Perspectives .

-...~-~.--.-- H TE01 20109 Under similar conditions both rat and mouse he- patic tissue could activate both 6-AC and the 3AI CSC to mutagenic intermediates. However, the pulmonary S-9 preparations were capable of ac- tivating aflatoxin B, (AFB,) (see Table 4). 7.8- dihydro-7.8-dihydroxybenaota)-8•diol pyrene-7. and 2-aminofluorene (2-AF) to mutagenic forms (Table 5). Thus. the lung preparations were metabolically Itceive by some measut+es, but did not activate 6-At'; or CSC. That the tiaslu hypotheti- cally at risk to tobacco-associsted carcinogenesis seems unable to activate the tobacco smoke con- densate allows many interesting interpretations which will be discussed in the last section of this report. In Vivo Studies Tobacco smoke condensate fractions have been shown to "promote" carcinogen-initiated skin tumors in mice (28. 29). Table 6 demonstrates that at least some fiacdons from the 2A1 CSC are capa- ble of acting synergistically with intrattac!teally in- stilled BP (with or without Fe,O,). resulting in a much higher incidence of Iung cancers in BC3FI mice: see Saft)ottP 130) for discussion on use of Fe=Q, in pulmonary cancer model systems. BP alaru (given every other week for 6 weeksl resulted in only one malignant tumor by 39 weeks after tteat- meot out of a total of 82 trr-ated animals (combining animals given 0.6 mg and 1.2 mg BP per treatment). TaEle 6. Actl.elm af T,8•d0ydno.7.84bldM4t+mWlolVy- 17$dnn _d 2.~SO~ I.LARI by p.lmra7 5•9• Pumunary S-9. ing tMOtein 2•AF. µg~tvbo 7.8dot. i.0'wbs AHH. v~+~ Survival. M~ 6.57 25 1901 d8 536 10 1925 81 668 10 1906 61 521 3.21 2J t4l2 32 458 10 142 52 540 5 1680 101 6Z7 0 23 - a 39 lo 86 52 6.l7 0.! 1928 37 SOA 3,28 0.3 If80 IB 338 0 1.0 - 88 76 0 0.1 - 99 86 TA 98 clone - - - 100 20 • Suspmswn assays pefornivd accoNlaB to pmceduma piren in Table 4. ' AHH - pnwtes 3.ON-BP fonmea per 33 mio iocubatlon in sepante aAee caominie8 23 ia8 BP/en as subasw. • Piticensep of vuble bacteria eRar 35 minuto assay nsorve to TA 9[I abre. TOW6B g.rtoatlsa ot ceew'd=pne en'Y=ar 2A1 CSC md itbctlDdhetftora ar ep ceodmmm=® BP.Infbtod 1®g ancan tn BQP, aseo.' Gel CSC t3aufon• Tmmmmt POUp selmee Bra Bib WA, N,w„ RF SM° No alber treatment 0/118 Oi37 1147 ty78 IM0 Or68 a'SO 0.6 ms FqQQ ON8 1147 071 m08 Or46 1J6J 0!SI 0.6 mg BP IM3 1lly6d fl7a 14159 14/82 4060 13138 1.2 m{ BP on9 9162 ]I]! 1Ld6 1N9 IOilB 9Y66 0.6 ei4 8P - mg Fq% 3160 7/61 a00 J40 ]I!0 3lu 11,36, 1.2 mj BP - 0.6 m8 Fqa 7169 9111 I Y77 ass I1P74 947 I yS0 ' Groops or BCpF,,C= fomale nvoe 18-1o wenlcs ald) wma mueuaeIeaDY 4TI inocdoted ooce eveay 2 weetu for n toml ota waoks wnb0.02 mt abdoi-al soimiom of BP. Fq0.. BP:Fqo, abas or In combimuom with the etpatte stooYe eondeeew ICSC) tfieatiom (10 µg eoahl. A< the end ot the "carciougm aeauoeae" peiod. tT treatment +itb the .ppropiam CSC Beotlom eree eontoreeA aoea ewey 2 weelu umi14 weelu WWto iaceiSee. aua ptisw++ud eeRveeatM ni mor larideete tor mke rnodomly meriBaO aRar 26 and 39 weetu on test. The tumor types dinQeu9eJ Wuopntlbtopeally foCluda nlveol4poie rdanowdnorean, edamiqunmau .;araioornne. puamwa aeU rarciuo®oe. unt uquamoae oaopfnme. A mom comlNen 4101161119111110111 of Ihnne lun8 mneen ie ym eieevheta li3). ' Dam Mvan in teims of the number of mice with luag carcinomas per the tomi number of oetmsU at risk. • RP ~ recommuosd ttacoons. ° S:A ~ Inrnn8 mniernel. C Apti11979 67

r H T®01 2ll 105 . Em•ir.,nmrntul Mruhh Persprrrnvs 1'.~l 29. pp. 6..i-0. 1979 D 'i®f®gical Activity of Tobacco Smoke and Tobacco Sm®ke-Related Chemicals by Richard E. Kourl,* Thomas H. Rude,* Rodger D. Curren,* Karen R. Brandt,* Ronald G. Sosnowskl,* Leonard M. Schachtman,* William F. Benedict,t and Carol J. Henry* Fspaaase to .Nolt eipmate smohe tlom ssfesenee cfliase:ttm cesntq in the pAmpllpeah s'ctlsy le 6 tusl. ha lbifty amtt 1- 2 tbildt. lodunlon or mmiot pulmomv~ mintisomal ~gtoese artl.a~. tbm satl.ity an be deoM[d by mdeg at str8nairoes eNYer (xnsaoipynstr or etGmtysawidln. and rao be Intdt>itsd by tnsotmmt .itA creloEalmfde or oetfnomydo D. Unntre tht lodaetfoo ar pa4ooeorr moooasy8tnnses rolbwiag intratrahe®1 adminhtratfoe st ].raatbyleholanthseat. Wme ef8uine smt:e4adooed Issdeass aam oot aeeqot.ecolly Iiaked t9 the Ah bavs. w[mte smake osedetmu a®d flactkoe desivad nam t6sw wodemta reat at Induae patmonarq moooosypmsin tatl.ut. el toe®N benmletlpeme mtmtawkm in vitro. al be metmhothed to fortm mametNe to 5afmonefla tpphimusnwn sestor stntles TA1338 ar TA98K dl trusdam C]R 10'[Sy tt8f in viho, tmd et eetsanee stm imeelingankity of 6tamtolpysem in morine pdmoeet7 ttsaos. A potantle0y Imposlam otmr.etloa tt tRS;I aleetees hepatk ttesae Is capmate of aelvaft .hele elgaem t®ohv rnadeamte to mwageubt fatms in ritro, murina pWmaoety Nsaoe does mt seem eaplbM oT saeh octl*a• don. Althoagh ttsese palmoeryrleti.ed daa. Aomogenma have sf;oYflmm AHtI estlritj, and am eeem0oft Aflaosta B.. iHSSnlsbtlnote+sa and 7.8~aa.8dlAydroartwasol®tpftaae to msdI , ro17nl. t4tie hemop68M! rCll to ®[nrlLQ both !ipitltee Sn10r14t tandenttR Y6d the QrWeIUBrO. 6• tsdssoe6ryt.ae. Thest twdls an dtetusaM vith refercaoe to tbe ewrcept that Nbete clgasem s'toote my be botb a potendta! .•lotdoW•.od ••psomxor'• of Img caneer m netee, and thm uele lolta popeetf euy Eti tta. mtal impottmt Is astermloing aeeer eitlt. . Introduction Cancer is not randomly distributed among the human population (1). ntther, both environmental (2) and genetic •tj-51 factora strongly influence its occurrence. Lung cancer is a specific example of a neoplasia whose occurrence is nonrandomly distributed. Its incidence is often associated with cigarette smoking (6). but the actual role that ta bacco smoke or smoke-related chemicals play in this association is unclear. A potential role of tobacco smoke involves in- teraction with those microsomally-bound mono- • NicTobsoloigiad Atsocistes. Depertment of Blochemieal On• :ology. rI River Road. Bethesda. Morvland _On16. * Chddn:n's Hospital af t.os Anteles. Los .ingeles. California vaoz° Apr11 1979 oxygenases that seem important in detetminina sus- ceptibility to chemically-induced cancer in model animal systems 17. 8). These enzymes are important because not only do they metabolize many en- vironmental chemicals. b'at also natulaUy occuning variations in their steady-state levels are genetically linked with susceptibility to toxicity. mutagenesis. and carcinogenesis induced by many of the same chemicals (7. 9-! l ). Three basic questions can be addressed con- cerqing the role of tobacco smoke in cancer suscep• tibility in ntan: (a) Does tobacco smoke contain chemicals that interact with the microsomal monooxygenases' (b) If they do interact, what at+e some oE the in i•mo and in ritnr efl'ects" 4cl Can certain individuals or tissues within an individual be at greater nsk than others from the effects of tobacco-related chemicals' 63

HTC012011 ( I 6 Hammond. E. C.. Garfinbel. L.. Seidman. IH.. and Lew. E. A. "Tar" and nuootine content of cigamtte smoke in rola- twn to death nues. Environ. Res. 12: 363 t 19761. 7. Kouri. R. E.. and Nebert. D. W. Genenc regulauon of sus- :eptibday to polyeyclie-hydrocatbon-mduad tumors in the mouse. In: Ongms of Human Cancer. CoId Spnng Harbor tabomtones Prese. Cold Spnng Harbor. N. Y.. 1977. 8. 77tmgrinson. S. S.. and Nebet•t. D. W. Genetic regulations of ihe metabolism of ehemrcal eammtgem and other foroign compounds. Adv. Cartecr Res. :3: 149 t 19771. 9. Koun. R. E.. Ratna. H.. Ill, and Whitmise. C. E. Genetic coeval of suscepubility to 3-methylcholamhsane•mduced subeuuuawue sareomas. Im. J. Cancer 13: 71a 11974). 10. Koun. R. E. Relattonship between levels of aryl hy- drocarbon hydroxyleae aetnuy and suseeptibility to 3• matlrylebolanthrone and bensatotpyrane-indecnd cancen in inbred strmns of mice. In: Polynuclear Aromatic Hy- droearbons: Chemistry. Metabolism and Ca,rcmogemtsts. R. 1. Fretdenthd and P. W. Jones. Eds.. Raven IYess. New York. 1916. p. 139. 11. Koun. R. E.. Rude. T. H.. Joglabar. R.. Dansette. P. M.. Jenma. D. M.. Attas. S. A.. Owens. 1. S.. and Nebert. 0. W. 2.3.7.&Tetrathlorodibrmzo-p-dioxin acts as cocan cinogen in causing 3•methylcholanthrene•iniuated sub- cutaneous tumors in mtee genetically "nomesponsive." at the Ab locus. Catxer Rea. 38: 2777 419781. 12. Koun. R. E.. Rude. T.. Thomas. P. E.. and Whitmire. C. E. Studies oo pulmonary aryl hydrocarbon hydtosylase activtty in inbved strains of mtce. Chem.-Biol. Interact. 13: 317 t19761. 13. Koun. R. E.. Denmise. C.. and Whnmme. C. E. The stgrtifi- cance of atyl hydrocarbon hydrosylase enzyme systems in the salectton of model svztems for respiratory car- nno®enests. In: Ext>f.nmental Lung Caneer. Caremogettests and B+oassays. E. Katbe and J. F. Parit. Eds.. Springer- VeriaB. New York. 1974. p. 48. 14. Van Centfott. J., and Gialen. J. 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M.. Jerina. D. N.. and Cooney. A. H. Mutagetticity and cytotoxiaty of benmlLtpytette arene ozedes. phenola. Quwnonee. and dihy. dtodiol6 in baemtid and meontoalian calls. Cancer Res. 36: 375011976y. 20. Fehmt. J. S.. and Nebatt. D. W. Mutagettssis of cmtam aa- dvtued catemo8res in .•Irro is associated with geneueally mRdlsted increases in monooxysettases atuvnY and oyto- chrome P,.450. J. Biol. Cbem. 2l0: 67691197l1. a. Neben. D. W.. Boobia. A. R.. Yagt. H.. Jetissa D. M.. and Kouns. R. E. Genatte dfRerenus in mouse eytoNnome P.- a50 etediated metabolism of benzaalpysene ln vinv aad cern cmogeoic index in rmu. In: Biologtcal Reactive Ioter• medota. D. J. Jo11ow. et a6. Eda.. PUmlm Ptees. N.w York. 1977. p. 125. ?:. Nebat. D. W.. Felton. l. S.. Kouri. R. E.. sed Boobie. A. R. Genetic dBfarneteea in the bindiag of beazotalpytene metabolites to DNA in the mouae. (n: Proeeediogti 3ni Inter- national Symposwm on Dtnection and Preoeadon of Csaar. H. E. Nieburqs. Ed.. Mmeel Dek(ser. 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H T 0 012010 I Does Tobacco Smoke Contain Chemicals Capable of Interacting with the RAonooxygenase Systems? Kinetics of increase of pulmonary AHH levels in C57BV6(661. DBAt2J t D2). and C3H1fCum iC31 strains of mice exposed to IAI cigarettes are shown in Figure I. Results suggested that there were rapid increases in pulmonary AHH activity that peaked by 6 hr post-exposure. and that these induced en- zymes remained at this level for at least 24 hr. The D2 strain was virtually nonresponsive to this smoke exposure compared with 56 and 0 strains; this re- sult paralleled that fsund when a polycyclic aro- matic hydrocarbon (PAH). 3-methylcholanthrene tMCI. was given intratracheally to the same strains (see Fig. l). However, the kinetics of induction of pulmonary AHH by smoke are different from those obtained when MC was used as an inducer. The peak of activity in the smoke induced lungs oc- curred at 6 hr (vs. 24 hr for MCI. Also the half-life of the smoke-induced enzyme was much longer (- 24 hr vs. - a hr). Finally, these smoke exposure condi- tions did not induce the AHH activity in liver. colon, and kidney tdata not shown) whereas MC• treatment did: monmver, filtered smoke (gas phasel did not induce enzyme activity in lung, liver, or FSount I. ?ultnonery response of C3WICum, C378L6Cum. and DBnIJ inl,nx! atteias of mxe to NC and whole cigarene intoke. Mice were enher uerltd with 200 pg MCM0.21°r gnlatin4aiitm solution inltatrecheafly t[T1 an described prevt- ously u21 and sacrifksd at the time periods specified or ex- poted to four cigarettes from a Walton smoking mschine: employing a 2•sec puff. 10% smoke. 30-sec expoeure. and 30-sec purge t111. Mice were given a.•min rest period before exposure to the next cigarette and were seetlfleed at varying ttmes after the founh cigarette. I.ungs were excised and frm- aen at -'0'C until assayed. AtIH units ate expressed as nmole 3-0tt-BP tornuNmmtg wet weight utisue t!,'1. .4ver- age acuvnr of thrae animals is given. 64 kidney tissue idata not ihownl. Smoke-induced AHH in the lung was similar to MC-induced AHH. in that both are capuble of ()-deethylation of ethoxyresorufin ITable I): see Burke and Mayer 1!: 1 for data on MC as an inducer. Data with ethoxyresorufin were particularly interesting be- cause studies have suggested that this chemical is a specific substrate for the P448 for P,-430) cyto- chrome (16). This is the cytochrome that is inti- mately associated with metabolizing ben- zo(aipyrene 1BP1 and MC to cytototic (17-19). mutagenic (2). DNA-binding 121. 22). and car- cinogenic (y. 10) forms. Similar to MC-induced AHH levels. the smoke-induced enzyme levels are dependent upon in viru protein and RNA synthesis Iree Table I). Although tobacco smoke induced AHH activity in the B6 strain preferentially to the D2 stntin. the genetic basis for this difference may not be the same as that for the difference in AHH activity induced by MC. Figure 2 shows that. whereas MC given intratracheally to (BI6D2) F, x D2 progeny induced pulmonary AHH activity in approximately 30% t 13/291 of the mice 02. 13). no clearcut discritnina- tion was observed in the smoke-exposed animels. However. it may be that the inducing capacity of cigarette smoke for pulmonary tissue is just too low to easily evaluate its genetic regulation. Whole tobacco smoke can be collected by con- densation with the use of Dry Ice. and this cigarette 3.,iv:.- ICSi:) or fractions derived therefrom 1231 can be evaluated for potential biological effects. The flactionation scheme and Table 1. rsRet of cyelotxaimlda and edlobfqyefn D an smedte. teelueed BC3F, tung ANH and stboxyrestiroft O40hyleea asL.tey: Treatment AHH O-Deethyiare activity. Bt:tivity ntttolaimm-g nntoldtmn-g lissYd tiaWC t+tacFane controls 0.33 <0.04 Smoke 0.70 0.61 Smoke - saline 0.67 0.83 SmDite - propylene glycol 0.33 0.83 Smoke - cyclohesrmde. 500 µtVg 0.32 014 Smoke - actinomycm D. I µgrg 0.42 0.15 ' Cyclohextmtde and actinomyein D were injected in- uaperiloneally t IPI in seline or in ptopytene gtyeol respectively. imtnedfately hafore exposure to one 2A 1 cigaretle on a lMatton stnolting machme. Medrods given in kgend to Fig. 1. Miee were sacriflced 6 hr atter tmoke exposure. Procedures were as de. scribed by Van Candon and Giden t1e1. • AHH unus are expressed as nmok 3-0H-BP foneed/mitvg wet weight of aesue. ,verage activity of three animals. ' U-tkethylase units are expressed as nmoles resorunn formeWmuug wet weight of tissue according to the procedures of Burlte and Mayer 1l51. Environmental Health Perspectives