Carcinogenic Polycyclic Hydrocarbons Research Articles

Analysis of technological conditions for the formation and burnout of carcinogenic pahs in boiler flue gases

THE PURPOSE. The necessity of establishing emission standards for highly toxic substances with carcinogenic properties in the context of the implementation of new principles of state regulation of environmental protection activities at energy enterprises is substantiated and confirmed. The analysis of the features of the formation and burnout of carcinogenic polycyclic aromatic hydrocarbons in the flue gases of boilers during the combustion of coals in lowtemperature conditions of the furnace process is carried out. Based on the analysis, the total and partial indicators of the harmfulness of flue gases were calculated in order to assess and predict the overall toxicity of coal combustion products, taking into account the contribution of carcinogenic substances.METHODS. When calculating the overall toxicity of flue gases, a systematic analysis and generalization of experimental data on the content of carcinogenic and non-carcinogenic polycyclic aromatic hydrocarbons in the combustion products of low-power boilers was applied.RESULTS. As a result of the performed study, the contribution of carcinogenic and non-carcinogenic polycyclic hydrocarbons to the overall toxicity of boiler flue gases during the combustion of Azeysky, Mugunsky, Cheremkhovsky and Tugnuy coal was determined. It is shown that substances such as benz(a)pyrene, fluoranthene, pyrene and phenanthrene significantly affect the level of the total indicator of the harmfulness of combustion products from 38.8 to 53.6%.CONCLUSION. The obtained research results can be applied at energy enterprises at the stage of substantiating the introduction of regime-technological and environmental measures in order to develop measures to improve environmental efficiency through technological rationing of carcinogenic emissions.

Comparative Study of the Properties of the Coal Extractive and Commercial Pitches

Three extractive pitches were produced by using thermal dissolution of the medium-ranked coals at 380 °C in anthracene oil. The empirical properties such as the chemical composition, solubility in quinoline and toluene, softening point, and content of carcinogenic polycyclic hydrocarbons, and also molecular and structural parameters of the extractive pitches were characterized in comparison with three commercially available pitches, including typical coal-tar pitch, petroleum-derived pitch, and blended pitch derived from the mixture of coal tar with petroleum feedstock. The molecular and structural properties of pitches were studied using FTIR, 1H and 13C NMR, and XRD techniques. It was shown that the average molecule of the extractive pitches was composed of predominantly aromatic rarely substituted pericondensed nuclei, such as that in coal-tar pitch. The spatial structures of both the extractive and reference pitches consisted of predominantly disordered carbon matter with a small amount of rather ordered nanosized “graphite-like” stacks. In terms of the technical specifications, the extractive pitches irrespective of the coal used met the requirements for the pitch binder and resembled commercial blended petro-coal-tar pitch. A remarkable merit of the extractive pitches compared to coal-tar pitch was low carcinogenicity (two to three times as low).

Carcinogen-specific mutational and epigenetic alterations in INK4A, INK4B and p53 tumour-suppressor genes drive induced senescence bypass in normal diploid mammalian cells

Immortalization (senescence bypass) is a critical rate-limiting step in the malignant transformation of mammalian somatic cells. Human cells must breach at least two distinct senescence barriers to permit unfettered clonal evolution during cancer development: (1) stress- or oncogene-induced premature senescence (SIPS/OIS), mediated via the p16-Rb and/or ARF-p53-p21 tumour-suppressive pathways, and (2) replicative senescence triggered by telomere shortening. In contrast, because their telomerase is constitutively active, cells from small rodents possess only the SIPS/OIS barrier, and are therefore useful for studying SIPS/OIS bypass in isolation. Dermal fibroblasts from the Syrian hamster (SHD cells) are exceptionally resistant to spontaneous SIPS bypass, but it can be readily induced following exposure to a wide range of chemical and physical carcinogens. Here we show that a spectrum of carcinogen-specific mutational and epigenetic alterations involving the INK4A (p16), p53 and INK4B (p15) genes are associated with induced SIPS bypass. With ionizing radiation, immortalization is invariably accompanied by efficient biallelic deletion of the complete INK4/CDKN2 locus. In comparison, SHD cells immortalized by the powerful polycyclic hydrocarbon carcinogen benzo(a)pyrene display transversion point mutations in the DNA-binding domain of p53 coupled with INK4 alterations such as loss of expression of p15. Epimutational silencing of p16 is the primary event associated with immortalization by nickel, a human non-genotoxic carcinogen. As SIPS/OIS bypass is a prerequisite for the immortalization of normal diploid human epithelial cells, our results with the SHD model will provide a basis for delineating combinations of key molecular changes underpinning this important event in human carcinogenesis.

Potent Modification of Inducible CYP1A1 Expression by Flavonoids

The present study examined modifications of β-naphthoflavone (β-NF)-induced cytochrome P450 1A1 (CYP1A1) expression by flavonoids in mouse hepatocytes in primary culture. Some flavonoids (apigenin, chrysin, flavone, flavanone, galangin, luteolin, and naringenin) by themselves induced CYP1A1 mRNA expression, especially flavone which was even more effective than β-NF. The effect on β-NF-induced CYP1A1 mRNA expression was varied, namely additive, suppressive, or both. An additive effect was observed after combined treatment with flavanone, naringenin, and chrysin, whereas kaempferol, myricetin, and quercetin decreased CYP1A1 levels. Apigenin, chrysin, galangin, luteolin, and morin synergistically enhanced β-NF-induced CYP1A1 expression at 24 h, but considerably suppressed it at 9 h. The structure-activity relationship of flavonoids affecting CYP1A1 expression as inducers or inhibitors is discussed. The present observations suggest the need to reveal the mechanism by which CYP1A1 expression is modified by flavonoids for risk assessment, since CYP1A1 activates environmental carcinogenic polycyclic hydrocarbons and flavonoids are major constituents in food.

Carcinogenicity of polycyclic hydrocarbons and their interaction with DNA

Carcinogenicity of polycyclic hydrocarbons and their interaction with DNA

Induction of Rad51 protein levels by p38 MAPK decreases cytotoxicity and mutagenicity in benzo[ a]pyrene-exposed human lung cancer cells

Rad51 is an essential component of the homologous recombination repair pathway. Abnormal expression of Rad51 has been reported in various carcinomas. Benzo[ a]pyrene (B[ a]P), a polycyclic hydrocarbon carcinogen found in the environment, induces cancer in multiple organs. B[ a]P has been shown to activate the p38 MAPK signaling pathway in mammalian cells. The prime purpose of this study was to determine how B[ a]P activates the p38 MAPK signaling pathway, and how this then regulates Rad51 expression in human cancer cells. Exposure of human lung cancer cells with B[ a]P increased Rad51 protein levels in a time- and dose-dependent fashion. B[ a]P also induced Rad51 mRNA and protein synthesis. Blockage of p38 MAPK activation by SB202190 or small interfering RNA (si-p38) decreased B[ a]P-elicited Rad51 protein levels by increasing Rad51 protein instability, but did not affect Rad51 mRNA transcription. Furthermore, enhancement of p38 MAPK signaling by constitutively active MKK6 (MKK6E) increased Rad51 protein levels and protein stability. Moreover, B[ a]P-induced cytotoxicity and mutagenicity were significantly increased in cells depleted of endogenous Rad51. Taken together, these results indicate that Rad51 protein provides a critical role in inhibiting the cytotoxicity and mutagenicity of B[ a]P in B[ a]P-treated human lung cancer cells. Furthermore, the work points to an unexpected role of p38 MAPK signaling in the control of Rad51 protein stability in response to B[ a]P exposure.

The role of repair protein Rad51 in synergistic cytotoxicity and mutagenicity induced by epidermal growth factor receptor inhibitor (Gefitinib, Iressa R) and benzo[ a]pyrene in human lung cancer

Rad51 protein is essential for homologous recombination repair of DNA damage, and is over-expressed in chemo- or radioresistant carcinomas. The polycyclic hydrocarbon carcinogen benzo[ a]pyrene (B[ a]P) affects MAPKs transduction pathways. Gefitinib (Iressa R, ZD1839) is a selective epidermal growth factor receptor tyrosine kinase inhibitor that blocks growth factor-mediated cell proliferation and ERK1/2 activation. We hypothesized that gefitinib enhances B[ a]P-mediated cytotoxicity by decreasing ERK1/2 activation. Exposure of human lung cancer cells to gefitinib decreased B[ a]P-elicited ERK1/2 activation and induced Rad51 protein expression. Gefitinib and B[ a]P co-treatment decreased Rad51 protein stability by triggering degradation via a 26S proteasome-dependent pathway. Expression of constitutive active MKK1/2 vectors (MKK1/2-CA) rescues the decreased ERK1/2 activity, and restores Rad51 protein level and stability under gefitinib and B[ a]P co-treatment. Gefitinib enhances B[ a]P-induced growth inhibition, cytotoxicity and mutagenicity. Co-treatment with gefitinib and B[ a]P can further inhibit cell growth significantly after depletion of endogenous Rad51 by siRad51 RNA transfection. Enhancement of ERK1/2 activation by MKK1-CA expression decrease B[ a]P- and gefitinib-induced cytotoxicity, and B[ a]P-induced mutagenicity. Rad51 protein protects lung cancer cells from synergistic cytotoxic and mutagenic effects induced by gefitinib and B[ a]P. Suppression of Rad51 protein expression may be a novel lung cancer therapeutic modality to overcome drug resistance to gefitinib.

Recombinant Enzymes Overexpressed in Bacteria Show Broad Catalytic Specificity of Human Cytochrome P450 2W1 and Limited Activity of Human Cytochrome P450 2S1

Human cytochromes P450 2S1 and 2W1 have received only limited attention with regard to characterization of function. Both cytochromes P450 have been reported to be overexpressed in human tumors, and cytochrome P450 2S1 is induced by carcinogenic polycyclic hydrocarbons. We report methods for high-level expression and purification of both cytochromes P450 from Escherichia coli, with the goal of establishing function. The level of expression of human cytochrome P450 2W1 achieved using codon optimization for E. coli was 1800 nmol of cytochrome P450 per liter of culture, the highest level achieved in this laboratory to date. Assays with a number of the typical cytochrome P450 substrates showed no detectable activity, including some for which qualitative reports have appeared in the literature. Cytochrome P450 2W1 catalyzed benzphetamine N-demethylation (k(cat), 3.8/min) and arachidonic acid oxidation, albeit at a very low rate (approximately 0.05/min). In a umu genotoxicity screen, cytochrome P450 2W1 catalyzed the activation of several procarcinogens, particularly polycyclic hydrocarbon diols, but cytochrome P450 2S1 did not. The bioactivation of procarcinogens by cytochrome P450 2W1 may be of significance in the context of reports of preferential expression of the enzyme in tumors, in that activation of procarcinogens could lead to the accumulation of mutations and enhance the carcinogenic process.

Differential requirement of signal pathways for benzo[ a ]pyrene (B[ a ]P)-induced nitric oxide synthase (iNOS) in rat esophageal epithelial cells

Overexpression of inducible nitric oxide synthase (iNOS) has been reported in several human cancers, including esophageal squamous cell carcinoma (SCC). Benzo[a]pyrene (B[a]P), a polycyclic hydrocarbon carcinogen found in tobacco smoke and in the environment, induces cancer in multiple organ sites in animals and may be a causative agent for certain human cancers, such as esophageal cancer. In the present study, the effects of B[a]P on the induction of iNOS and the signaling pathways that lead to the induction were investigated in cultured rat esophageal epithelial (RE-149) cells. Treatment of RE-149 cells with B[a]P led to a marked increase in the expression of iNOS. The induction of iNOS by B[a]P was found to occur through an extracellular signal-regulated protein kinases (ERKs)-dependent pathway, since inhibition of ERKs by either pretreatment of RE-149 cells with PD98059, an inhibitor of ERKs upstream kinase MEK1/2, or overexpression of DN-ERK2, blocked the induction of iNOS by B[a]P. Furthermore, impairing nuclear factor-kappaB (NFkappaB) activation by either NEMO-BDBP, an NFkappaB specific inhibitor, or overexpression of DN-IkappaBalpha or IKK-KM markedly inhibited the expression of B[a]P-induced iNOS, suggesting that the NFkappaB pathway is also required for the induction of iNOS by B[a]P. In addition, treatment of RE-149 cells with either SB202190, a p38 kinase inhibitor, or c-JunN-terminal kinase inhibitor II, resulted in an increased induction of iNOS. Pretreatment of RE-149 cells with wortmannin, a PI-3K inhibitor, or with rapamycin, an mTOR/p70S6K pathway inhibitor, had no effect on the expression of iNOS. These results suggest that B[a]P initiates the signaling pathways leading to the induction of iNOS in cultured rat esophageal epithelial cells. In view of the potential role of iNOS in the development of esophageal SCC in humans, we speculate that the induction of iNOS by B[a]P may be one mechanism by which B[a]P could produce carcinogenic effects in the human esophagus.

Impact of p53-based immunization on primary chemically-induced tumors

In mice as well as humans, cytotoxic T lymphocytes (CTL) specific for wild-type-sequence (wt) p53 peptides have been shown to react against a wide range of tumors, but not normal cells. As such, they are attractive candidates for developing broadly applicable cancer vaccines. Of particular interest is the potential of using p53-based vaccines in high-risk individuals to prevent cancer. Methylcholanthrene, an immunosuppressive polycyclic hydrocarbon carcinogen implicated as a causative agent in human cancers, has long been used to induce murine tumors with a high incidence of genetic alterations and sensitivity to wt p53-specific CTL. To analyze the potential of p53-based vaccines on primary tumors, we evaluated the efficacy of DNA and dendritic cell vaccines targeting wt p53 peptides given to methylcholanthrene-treated mice in the protection or therapy settings. The results indicate that the efficacy of these vaccines relative to reducing tumor incidence were severely compromised by vaccine-induced tumor escape. As compared to tumors induced in non-immunized mice, a higher incidence of epitope-loss tumors was detected in tumors from the immunized mice. The increase in tumor escape arose as a consequence of either increased frequencies of mutations within/flanking p53 epitope-coding regions or downregulation of expression of the major histocompatibility complex Class I molecules that present these epitopes for T cell recognition These findings are consistent with current views of immunoselection occurring in patients receiving tumor peptide-based immunotherapy, and impact on the design and implementation of p53-based vaccines, in particular, those aimed at treating individuals at high risk for developing cancer.

Modulation of Carcinogen Metabolism and DNA Interaction by Calcium Glucarate in Mouse Skin

Almost all the polycyclic aromatic hydrocarbons (PAHs) require metabolic activation to exert their carcinogenic activity. Environmental carcinogen [(3)H] benzo[a]pyrene (BP) is carcinogenic only after its metabolic transformation to a reactive intermediate, which can then bind to cellular macromolecules. Inhibition of dimethylbenz anthracene- (DMBA-) DNA binding generally accompanied inhibition of tumor initiation as most inhibitors of initiation interfere with the metabolic activation of the initiator. The importance of carcinogen-DNA interaction and the enzymes involved in the metabolism of carcinogenic polycyclic hydrocarbons has led to a search for inhibitors that would be useful in modifying the cancer-causing effects of the PAHs. We tested the effect of calcium glucarate (Cag), a naturally occurring nontoxic compound, on carcinogen metabolism and DNA interaction. Cag inhibited [(3)H] BP binding to both calf thymus DNA in vitro and to epidermal DNA in vivo. Application of Cag to mouse skin caused a dose-dependent inhibition of [(3)H] BP binding to epidermal DNA. To establish the relevance of the in vivo results to the in vitro situation, we followed the in vitro effect of Cag on [(3)H] BP binding to calf thymus DNA and observed that Cag inhibited the [(3)H] BP binding to calf thymus DNA in the presence of microsomes prepared from animals treated with DMBA. We also studied related events like DNA synthesis and carcinogen metabolism. For assessing the DNA synthesis, thymidine kinase was used as marker. Cag caused a dose-dependent inhibition of DMBA-induced thymidine kinase activity. At the same time, Cag caused a marked inhibition of DMBA-induced aryl hydrocarbon hydroxylase (AHH) activity, an enzyme responsible for the metabolism of PAHs like BP, both in vivo and in vitro. Our study indicates that Cag exerted its antitumor effect possibly by inhibiting the carcinogen-DNA binding, which appears to be due to reduced DNA synthesis and AHH activity.

Lésions provoquées chez le criquet Gomphocerus Sibiricus par l'absorption de benzo- a-pyrène et de 3-méthylcholanthrène

Using UV light in order to study accumulation of BaP or 3MC by Gomphocerus sibiricus, a locust strictly located between 2 000 and 2 800 m, we have previously shown that these HAP can be rapidly absorbed by midgut, then, accumulated in many organs and essentially eliminated by Malpighian tubules (J.B., 1999), presumably after partial degradation. Now we describe the effects of these carcinogenic polycyclic hydrocarbons at histological and cytological levels. Wholly invaded, midgut epithelium is profoundly intoxicated, numerous cells showing structural abnormalities or beeing destroyed: excessive merocrine secretion becoming finally holocrine. Connective and muscular tissues do not constitute effective barriers against toxic effects of the HAP. Fat body and muscles, though strongly impregnated with each of them, show only discrete cellular damage. Locally, the reproductive system is more clearly affected. Otherwise, nervous cord ganglia contain some dead cells, and we describe an effective swelling of the intercellular spaces. This point seems to imply disturbancies of the glial sheats which isolate neurons from their neighbours. Finally, digestive caeca, strongly affected by HAP, are partly destroyed, especially after treatment by BaP. We suggest caeca could be a secondary route of degradation and excretion of HAP. Tubes of Malpighi seem to be the principal route of their elimination; their epithelium is frequently severely injured and may sometimes disappear. As with mammals, we have to underline here similar differential cell susceptibilities within each organ of a locust. However what may be, we have perceived effective disturbancies which are strong enough to involve a knock-down and finally death of a part of the treated locusts. This work gives us new opportunities for ulterior research, for example, with locusts sampled in a variety of biotopes, strongly polluted or “not”. Our UV test may be used as a first approach.

A Novel DNA Damage Checkpoint Involving Post-transcriptional Regulation of Cyclin A Expression

The intracellular metabolism of many carcinogenic polycyclic aryl hydrocarbons (PAHs, typified by the ubiquitous pollutant benzo[a]pyrene or B[a]P) generates electrophilic products that react covalently with genomic DNA. Cells that acquire PAH-induced DNA damage undergo growth arrest in a p53-independent manner (Vaziri, C., and Faller, D. V. (1997) J. Biol. Chem. 272, 2762-2769). In this report we have investigated the molecular basis of PAH-induced cell cycle arrest. Mitogenic signaling events involving cyclins D and E, Rb phosphorylation, and transcriptional activation of E2F-responsive genes (including cyclin E and cyclin A) were unaffected in cells containing PAH-damaged DNA. However, PAH-induced growth arrest was associated with post-transcriptional decreases in cyclin A expression. Mitogen-induced expression of cyclin B, an event that is temporally distal to cyclin A expression, was also inhibited in PAH-treated cells. The PAH-induced cell cycle block was transient, and arrested cells resumed DNA synthesis after a prolonged ( approximately 20 h) delay. Resumption of DNA synthesis in PAH-treated cells occurred concomitant with elevated expression of cyclins A and B. PAH-induced cell cycle arrest was overcome by ectopically expressed cyclin A (encoded by a recombinant adenovirus in transiently infected cells). Overall, our results suggest the existence of a DNA damage checkpoint pathway that arrests cell cycle progression via post-transcriptional control of cyclin A expression.

The effects of carcinogenic methylcholanthrene on carbohydrate residues of NK cells.

The present study examines the effect of methylcholanthrene (MCA), a a carcinogenic polycyclic hydrocarbon, on the carbohydrate receptor determinants (RD) on natural killer (NK) cell surface using the bead-coupled lectin assay. Murine NK cells exhibited different degrees of preferential binding to the specific lectins tested. Of the ten lectins tested, five exhibited a positive binding affinity while the remaining five exhibited no or insignificant binding. NK cells bind to beads derivatized with mannose specific lectins: Concanavalin A (Con A), Lens culinaris, and Pisum sativum. NK cells also bind to other lectin beads such as Triticum vulgaris (GalNac) and Vicia villosa (D-GlcNAc). All these lectin beads exhibited greater than 90% adhesion. The underivatized control beads exhibited no NK binding. The NK cells that were exposed to MCA for 2 h demonstrated a significant decrease in lectin bead-cell coupling in a dose dependent manner. MCA (10 micrograms/mL) caused a 17.8%, 40% and 4.7% decrease in binding affinity when introduced to the mannose specific lectins; Con A, L. culinaris and P. sativum beads, respectively. The binding of T. vulgaris and V. villosa to NK cells was inhibited (23.4% and 28%) by MCA treatment. An increase in the dose to 20 micrograms/mL resulted in a greater inhibition in binding affinity towards lectin beads. Con A, 35.3%, L. culinaris, 62.6%, P. sativum, 30.9%, T. vulgaris, 44.2% and V. villosa, 46.2%. The effect of MCA activation and cytotoxic response. Hydrolysis of PI metabolites (PIP and PIP2) cause generation of secondary messenger: inositol-1,4,5-triphosphate and diacylglycerol, both of which elicit an immune response through their products (Ca2+ and PKC) respectively. Identification of the relationship between receptor level, induction of second messenger and cytotoxic activity may resolve the molecular basis of suppression of NK cytotoxicity by MCA and other PAH compounds.

Diallyl sulfide down-regulates polycyclic aromatic hydrocarbon-induced cytochrome P450 1A1 in mouse liver and lung

The expression of cytochrome P450 genes directly within target cells is an important determinant of human susceptibility to cancers and other chemically initiated diseases. One pivotal gene, C Y P 1 A 1 codes for an inducible cytochrome P450 isozyme 1A1 responsible for the bioactivation of numerous carcinogenic polycyclic hydrocarbons. In the present study, the effects of 3-methylcholanthrene, a polycyclic aromatic hydrocarbon, on the activity and expression of CYP1A1 and the protective effects of diallyl sulfide on 3-methylcholanthrene-induced changes in mice liver and lung were investigated. After a four daily 3-methylcholanthrene-treatment (25 mg/kg, i.p.), liver and lung microsomal 7-ethoxyresorufin-Odeethylase (EROD) activity, associated with CYP1A1, was increased. A corresponding increase in the level of CYP1A1 mRNA was observed in mouse liver and lung after 3-methylcholanthrenetreatment by Northern blot analysis. Diallyl sulfide reduced 3-methylcholanthrene-induced CYP1A1 mRNA expression and its associated EROD activity in mouse liver and lung. The modulation of CYP1A1 mRNA by 3-methylcholanthrene and diallyl sulfide was mainly due to transcriptional regulation.

The mutagenicity of dibenz[a,j]acridine, some metabolites and other derivatives in bacteria and mammalian cells.

Dibenz[a,j]acridine (DBAJAC) was studied because of its close structural relationship with a number of important carcinogenic polycyclic and azaaromatic hydrocarbons. It was of particular relevance to examine the mutagenicity of known or proposed 'bay-region' metabolites, which may be proximate or ultimate carcinogenic derivatives of DBAJAC. Trans-1,2-, 3,4- and 5,6-dihydrodiols, the 4- and 6-phenols, the 5,6-oxide and N-oxide derivatives, and anti- and syn-3,4-diol 1,2-epoxides of DBAJAC were examined for their mutagenicity in Salmonella typhimurium TA98 and TA100 and in V79 Chinese hamster lung cells. Of all the compounds studied which require metabolic activation, the 3,4-dihydrodiol was the most active in both TA100 and in V79 cells. The activity of the 3,4-dihydrodiol enantiomers was also tested in strain TA100 where no difference was observed from that of the racemic mixture. In V79 cells only the 3R,4R-dihydrodiol was active, the activity being about three times that of the racemic material. Salmonella strains TA98 and TA100 also differed in their sensitivity towards DBAJAC dihydrodiols, the 1,2-isomer being of greatest activity in TA98. The most mutagenic compounds in both mammalian and bacterial cells were the 'bay-region' diol epoxides of DBAJAC which did not require metabolic activation by S9 mix. The anti-DBAJAC 3,4-diol 1,2-epoxide was more mutagenic than the syn form in V79, TA98 and TA100 cells. Overall these results suggest that the in vivo biological activity of DBAJAC metabolites is likely to reflect previous findings with other similar polycyclic aromatic hydrocarbons.

Heterogeneity of biotransformation of fluoranthene in perifused liver cells.

Pathways of biotransformation of the carcinogenic polycyclic hydrocarbon fluoranthene in individual isolated perifused liver cells were delineated using noninvasive microfluorescence spectroscopic techniques. An example of heterogeneity of coupling between phase I and phase II enzymes detected in a population of beta-naphthoflavone-induced rat liver cells demonstrates the usefulness of the experimental model of perifused liver cells.

Induction of genetic tandem duplications in Salmonella by polycyclic aromatic hydrocarbon and amine carcinogens

6 polycyclic aromatic hydrocarbon or similar amine carcinogens were tested as inducers of genetic tandem duplications in a rough strain of Salmonella typhimurium. When metabolically activated by rat-liver microsomes, all 6 were active in inducing genetic tandem duplications, yielding from over 3 times to almost 14 times as many tandem duplicants per viable bacterium as did concurrent uninduced control cultures. These results extend the number and chemical diversity of carcinogens shown to induce genetic duplications in bacterial tester systems. We suggest that polycyclic hydrocarbon carcinogens may act in carcinogenesis by inducing genetic duplications or other genetic rearrangements. Duplication induction may be a useful genetic endpoint for screening potential carcinogens.

Enhanced formation of benzo(a)pyrene: DNA adducts in monocytes of patients with a presumed predisposition to lung cancer

Blood monocytes from 45 selected patients with lung cancer and 30 healthy controls were incubated with [G-3H]-benzo(a)pyrene for 30 h, and the formation of covalently bound DNA adducts was determined. The lung cancer patients were either relatively young (below 46 yr), nonsmokers, or had at least one first degree relative with lung cancer. Therefore, they might be considered cancer prone. The DNA adducts were significantly elevated in 22 patients with early age cancer (4.34 fmol/micrograms of DNA; P less than 0.04). In 12 familial cases, the slight elevation (2.77 fmol/micrograms of DNA) was not statistically significant in comparison to healthy controls. Benzo(a)pyrene:DNA adduct levels did not differ significantly between smokers and nonsmokers. Eight of 9 lung cancer patients with DNA adducts equal or above 4.5 fmol/micrograms of DNA but only 16 of 36 with adducts below this value had either oat cell or squamous cell cancer (P less than 0.05). The observed enhanced formation of covalent DNA adducts in blood monocytes exposed to a carcinogenic polycyclic hydrocarbon may be genetically determined and could play a role in the development of lung cancer at an early age.

Virus-like particles in cultured C3H/St mouse cells treated with a carcinogenic polycyclic hydrocarbon.

Virus-like particles with cylindrical form were found in cultured alveolar macrophages and lung fibroblasts of C3H/St mice, after treating these cells with a carcinogenic polycyclic hydrocarbon, 1, 2, 5, 6,-Dibenzanthracene (DBA). Their morphology looked identical to those observed in vivo in the reactive cells which engulfed DBA crystals implanted into the brain and muscle of the same mouse strain. These particles were not found in either the untreated cells of C3 H/St mice or the treated cells of BALB/c mice. In the alveolar macrophages, these particles appeared first at 3 days after DBA treatment and reached the maximum number around the 30th day. They still kept their morphology in the degenerating cells which had lost the cytoplasmic organelles. These findings suggest the possibility that DBA induced the expression of the viral genome endogeneous to C3H/St cell.