Decreased Expression of Cytochrome P450 mRNAs and Related Steroid Hydroxylation Activities in Hepatic Hyperplastic Nodules in Male F344 Rats

Abstract

To explore the mechanism(s) underlying the relationship between expression of cytochrome P450 (CYP) enzymes and growth of hepatic hyperplastic nodules, mRNA of CYP1A1, 1A2, 2B1, 2B2, 2C6, 2C11, and 3A1, and CYP enzyme-mediated hydroxylation of testosterone (T), I 7β-estradiol (E), and corticosterone (C) were determined in a group of selected large nodules from livers of male F344 rats given the modified Solt-Farber resistance protocol and compared with control and surrounding tissue. Slot-blot analysis with specific oligonucleotides showed little or no mRNA of CYP2B1, 2B2, and 2C6 in nodules, surrounding tissue, and control liver without PB treatment. Constitutive expression, as many endogenous CYP1A2, 2C11, and 3A1 in nodules was from 2 to 10 times less in nodules than in control liver. PB treatment increased mRNA of CYP21, 2B2, 2C6, 2X11, and 3A1 in control and surrounding tissue substantially, but was 3 to 11 times less in the nodules than in the control liver. Messenger RNA of CYP1A1 was not detected in either nodules or control livers independent of PB treatment. HPLC determination demonstrated a general pattern of a decrease on CYP enzyme-mediated hydroxylation of T, E, and C in the nodules relative to the control liver. The differences between nodules and control liver were usually 3 to 10-fold for the 12 metabolites detected although smaller decreases were observed for a few metabolites in some nodules. PB increased the rate of hydroxylation for 11 of the 12 metabolites and resulted in 2 additional metabolites of both T and E in control liver and some nodules. However, the general pattern of lower hydroxylation activities in the nodules relative to the surrounding tissue remained the same as that in the non-PB group. These observations are consistent with the hypothesis that intracellular kinetics of endogenous CYP enzyme substrates may differ in some nodules, relative to the surrounding tissue, as a result of decreased expression of CYP genes. This difference in CYP expression pattern in turn may contribute to the selective growth and progression to cancer of certain nodules, as many endogenous CYP enzyme substrates are modulators of DNA and RNA synthesis and cellular growth and differentiation.