Cx32 is a major gap junction protein of the liver and is often aberrantly expressed in liver tumours. We have studied mutation of the Cx32 gene during chemically induced hepatocarcinogenesis. DNA from 12 rat liver tumours induced by diethylnitrosamine or N-ethyl-N-hydroxyethylnitrosamine (EHEN) was analysed by the PCR/SSCP method. One tumour induced by EHEN harboured a G--> A transition mutation at codon 220, substituting His for Arg. When the mutant DNA was transfected into HeLa cells, which are deficient in gap junctional intercellular communication (GJIC), GJIC recovered, as in HeLa cells transfected with the wild-type Cx32 gene. Moreover, GJIC was modulated by cAMP, 12-O-tetradecanoylphorbol-13-acetate and lysophosphatidic acid similarly in mutant and wild-type Cx32 transfectants. These results suggest that Cx32 gene mutations are rarely involved in rat hepatocarcinogenesis and that the mutation found in a tumour may be functionally silent.

The reactions of benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene metabolites and of r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene with the DNA of matrix-bound and released chromatin fractions of rat-liver nuclei have been examined. Qualitatively there were no differences between the DNA-bound metabolites in each fraction but more binding to matrix-bound DNA occurred. Evidence was obtained that the increased binding of hydrocarbon to matrix-bound DNA was not dependent upon the proximity of hydrocarbon-metabolizing enzymes and Sephadex LH20 chromatography showed that the differences between the fractions were not due to contamination of DNA with residual proteins. The conformation of the matrix-bound chromatin may make its DNA more accessible to reactive metabolites than that of released chromatin.

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