W. M. Bracken

Skin tumor initiating ability of benzo(a)pyrene 4,5- 7,8- and 7,8-diol-9,10-epoxides and 7,8-diol

The skin tumor initiating abilities of both K-region and non-K-region epoxides of benzo(a)pyrene(BP) were determined in mice using a two-stage system of tumorigenesis. BP-4,5-epoxide and BP-7,8-dihydrodiol-9,10-epoxide (anti) were found to be weak tumor initiators whereas BP-7,8-epoxide had about a third of the activity as the parent hydrocarbon, BP. However, the 7,8-dihydrodiol of BP was found to be approximately as potent as BP suggesting that it may be a proximate carcinogen.

The effects of weak or non-carcinogenic polycyclic hydrocarbons on 7, 12-dimethylbenz[a]anthracene and benzo[a]pyrene skin tumor-initiation*

Benzo[e]pyrene (B[e]P) inhibited 7,12-dimethylbenz[a]anthracene (DMBA) skin tumor-initiation in mice by 84%, whereas pyrene and fluoranthene inhibited DMBA initiation by 50 and 34%, respectively. However, B[e]P, pyrene and fluoranthene had either no significant effect or a slight enhancing effect on benzo[a]pyrene (B[a]P) skin tumor-initiation. In addition, B[e]P had essentially no effect on the initiating ability of (+/-)B[a]P-7 beta,8 alpha-diol-9 alpha,10 alpha-epoxide. As a tumor-initiator, B[e]P was found to have very weak activity at a 252 microgram/level (0.4 papillomas/mouse at 40 weeks) and no activity at 100 microgram. When given at a dose of 100 microgram twice weekly, B[e]P induced 2.1 papillomas/mouse at 30 weeks, and 25% of the mice had carcinomas at 40 weeks. However, B[e]P carcinogenic activity is weak when compared to B[a]P, which can induce a comparable tumor response at a dose of 5 microgram twice weekly. When B[e]P was tested as a tumor promoter at a dose of 100 microgram twice weekly after DMBA initiation, it induced 4.5 papillomas/mouse at 30 weeks and a 45% carcinoma incidence at 40 weeks, which was approximately twice as effective as B[e]P alone. The data show that B[e]P is a very weak tumor initiator, a weak complete carcinogen, a moderate tumor promoter, possibly a weak co-tumor-initiator when given with B[a]P, and a potent anit-tumor-initiator when given with DMBA. The anti-tumor initiating and co-tumor-initiating effects of B[e]P appear to be related to its ability to modify the conversion of the tumor initiator into an electrophilic intermediate(s) which are capable of covalently binding to DNA. In addition, B[e]P induced epidermal cellular proliferation which may be related to its promoting ability.

The ability of enteric bacteria to catalyze the covalent binding of bile acids and cholesterol to DNA and their inability to metabolize benzo(a)pyrene to a binding product and to known metabolites

The capacity of enteric bacteria (E. coli, Salmonella, Pseudomonas, Shigella and Klebsiella) to catalyze the covalent binding of benzo(a)pyrene (BP), cholic acid, deoxycholic acid and cholesterol was investigated. In general, these bacteria were incapable of activating BP to a covalently bound product with calf thymus DNA. Metabolism studies of BP by fluorometric assay failed to indicate any accumulation of BP-3-hydroxy in the incubation medium. Detailed metabolic investigation with high-pressure liquid chromatography indicated that these bacteria did not produce any known metabolites which are formed by mammalian systems. However, radioactivity was detected in all fractions, suggesting that the bacteria were readily metabolizing BP into smaller molecules for energy and carbon sources. Although the enteric bacteria did not metabolize BP into known metabolites, some were capable of activating cholesterol, cholic acid and deoxycholic acid to covalently bound products with DNA. The binding data with cholesterol and bile acids also suggested that the binding process required NADPH as a cofactor because binding level was rather low without NADPH.