Analysis of c-myc expression in a human hepatoma cell line

A tumorigenic human hepatoma cell line, Hep G2, has been shown to have high steady-state levels of c-myc transcripts compared to normal human liver. We have now characterized c-myc expression in Hep G2 cells with regard to message stability, gene rearrangements, gene amplification, chromosomal translocations, promoter utilization, and the effects of protein synthesis inhibitors. We have determined that the half-life of the Hep G2 c-myc transcript is approximately 20 min and conclude that the high steady-state level of c-myc mRNA is not the result of a specific stabilization of the c-myc message but probably results from increased c-myc gene transcription. c-myc expression in Hep G2 cells appears to be constitutive, since it remains constant in different cell growth states (log phase versus nondividing cells). The high constitutive expression of the c-myc gene in Hep G2 cells could not be explained by gene amplification, gene rearrangements, or chromosomal translocations. However, based on an S1 nuclease protection assay, the P1/P2 promoter utilization ratio is approximately 1/1 which differs from the 1/5 P1/P2 ratio observed in normal human liver. Treatment with cycloheximide, a protein synthesis inhibitor, does not superinduce Hep G2 c-myc transcription based on transcription "run on" and RNA slot blot analysis. However, cycloheximide treatment does exert a posttranscriptional effect involving the specific stabilization of the c-myc message.