INSULIN PROMOTES THE BIOSYNTHESIS AND SECRETION OF APOLIPOPROTEIN B-48 BY ALTERING APOLIPOPROTEIN-B MESSENGER-RNA EDITING

Long-term insulin treatment selectively stimulates secretion of the tr uncated form of apolipoprotein B (apoB), apoB-48, from primary rat hep atocytes in culture. Chronic treatment with insulin at 400 ng/ml cause s a 3-fold increase in total apoB secretion, with apoB-48 making up ab out 75% of that increase. apoB-48 is the protein product generated by translation of full-length apoB mRNA which has been modified by a post transcriptional editing mechanism. Editing changes codon 2153 in the m iddle of the apoB-100 coding region from CAA, coding for glutamine, to UAA, a translation stop signal. We therefore examined the effect of i nsulin treatment on the ratio of edited to nonedited apoB mRNA in RNA isolated from primary rat hepatocyte cultures. There was a dramatic sh ift in the ratio of edited versus nonedited forms of apoB mRNA, from a bout 1:1 in untreated cells to 7:1 in insulin-treated cells. Insulin e xerted a dose-dependent effect on apoB secretion and apoB mRNA editing over the range of insulin concentrations studied (0.4-400 ng/ml). In contrast, oleic acid, which also increased apoB (B-48 and B-100) secre tion, had no significant effect on the ratio of apoB-48 to apoB-100 pa rticles secreted and no effect on the proportion of edited apoB mRNA. Neither insulin nor oleic acid affects total apoB mRNA levels as assay ed by Northern blot analysis. These data strongly suggest that insulin stimulates biosynthesis and secretion of apoB-48 in rat hepatocytes b y regulating the proportion of edited apoB mRNA.