TRANSPLACENTAL TRANSFER OF POLYCHLORINATED-BIPHENYLS INDUCES SIMULTANEOUSLY THE EXPRESSION OF P450 ISOENZYMES AND THE PROTOONCOGENES C-HA-RAS AND C-RAF

At day 15 of gestation, rats were injected with a single i.p. dose of 100, 250 and 500 mg/kg body weight of a mixture of polychlorinated biphenyls (PCBs) (Aroclor 1254). Seven days later, significant increases in maternal and foetal cytochrome P450, cytochrome b5 and cytochrome c (P450) reductase were found. Concomitantly, the metabolism of nitroanisole, aniline, ethoxyresorufin and benzo[a]pyrene was significantly increased, but foetal metabolism of dimethylnitrosamine was not detectable and only marginal increases in the metabolism of aminopyrine and aldrin were seen. In contrast, maternal metabolism of dimethylnitrosamine, aminopyrine and aldrin was measurable, but significant increases were determined only with the latter substrate. Transplacental transfer of PCBs resulted in increased metabolism of substrates catalysed by foetal CYP1A1 and CYP2B1, but there was no evidence for CYP2E1-catalysed reactions. Further measurements show significant increases in foetal and maternal epoxide hydrolase, glutathione-S-transferase and UDP-glucuronyl transferase activities, thus suggesting that treatment with Aroclor 1254 resulted in coordinated increases in foetal and maternal oxidative and post-oxidative drug metabolism. Western blot analysis of microsomal proteins shows the induction of foetal and maternal CYP1A1, CYP1A2, CYP2A1, CYP2B1, CYP3A1 and CYP4A1. In addition, increased expression of CYP2C6 was seen with the mother but not the foetus. Unlike the mother, foetal rats did not express CYP2E1 and the expression of the above-listed P450 isoenzymes was greater in the mother than the foetus. Northern blot analysis shows significant increases in maternal and foetal CYP1A1, CYP1A2 and CYP2B1 mRNA. An increased amount of CYP3A1 mRNA was only seen with the mother, but not the foetus. Treatment of mothers with Aroclor 1254 resulted in reduced CYP2A1, CYP2C7, CYP2E1 and CYP4A1 mRNA. Insignificant differences in the expression of foetal CYP2A1 and CYP4A1 mRNA were found, but in utero exposure to PCBs reduced the amounts of CYP2E1 mRNA and there was no foetal CYP2C7 mRNA transcript. Treatment with Aroclor 1254 increased the expression of the protooncogenes c-Ha-ras and c-raf in the mother and the foetus, but at varying intensities. Pregnancy itself was linked to an increased expression of these protooncogenes. erbA and erbB mRNA was not detected.