Preferential effects of the chemotherapeutic DNA crosslinking agent mitomycin C on inducible gene expression in vivo.
The immediate effects of a single dose of the chemotherapeutic DNA crosslinking agent, mitomycin C (MMC), on the expression of several constitutive and drug-inducible genes were examined in a simple in vivo system, the 14 day chick embryo. We observed no effect of MMC on the steady-state mRNA expression of the constitutively expressed beta-actin, transferrin, or albumin genes. In contrast, MMC treatment significantly altered both the basal and drug-inducible mRNA expression of two glutethimide-inducible genes, 5-aminolevulinic acid (ALA) synthase and cytochrome P450 CYP2H1. The basal expression of these genes was transiently but significantly increased over a 24 hr period following a single dose of MMC. Conversely, MMC significantly suppressed the glutethimide-inducible expression of these genes when administered 1 to 24 hr prior to the inducing drug. The effects of MMC on both basal and drug-inducible ALA synthase and CYP2H1 mRNA expression were principally a result of changes in the transcription rates of these genes. In contrast, MMC treatment had little or no effect on glutethimide-induced expression of ALA synthase or CYP2H1 when administered 1 hr after the inducing drug, suggesting that a very early event in the induction process represents the target for these MMC effects. Covalent binding studies demonstrated that the effects of MMC on gene expression were closely correlated temporally with formation of [3H]-porfiromycin-DNA adducts. These results support the hypothesis that genotoxic chemicals specifically target their effects to inducible genes in vivo.
Caron
R M
RM
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, NH 03755-3835.
Hamilton
J W
JW
D000624:5-Aminolevulinate Synthetase / Q000235:genetics; D000818:Animals; D002642:Chick Embryo; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D008099:Liver / Q000201:enzymology; D016685:Mitomycin / Q000494:pharmacology*; D011160:Porfiromycin / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics
D003432:Cross-Linking Reagents; D012333:RNA, Messenger; D016685:Mitomycin; D003577:Cytochrome P-450 Enzyme System; D000624:5-Aminolevulinate Synthetase; D011160:Porfiromycin