The level of expression of the rat growth hormone gene in liver tumor cells is at least eight orders of magnitude less than that in anterior pituitary cells

Abstract

Rat liver hepatoma cells (HTC) which express liver-specific gene products were assayed for the expression of the rat growth hormone (rGH) gene, which is normally expressed in anterior pituitary somatotrophs. The combination of immunoprecipitation and two-dimensional gel electrophoresis provided a highly sensitive assay for rGH synthesis at levels as low as one part in 10(9) of cell protein synthesis (or four molecules of rGH per cell). No rGH expression was detected at this level. The lack of expression in HTC cells did not derive from a deletion of the rGH gene, as shown by Southern hybridization analysis of genomic DNA. Because the gene is expressed at greater than 30% of anterior pituitary protein synthesis, differentiation regulated rGH expression by over 10(8)-fold between the two cell types. Additionally, DNA-excess solution hybridization was used to measure the level of rGH mRNA sequences. A novel and general method for preparing single-strand probes from recombinant plasmids was developed. Hybridization analyses with a sensitivity of detection of 1 part in 10(8) failed to detect any rGH RNA sequences in either the nucleus or cytoplasm of HTC cells. It is concluded, therefore, that the restriction in rGH expression in the liver tumor cells is likely to occur at the level of the transcription of the gene, and that for all practical purposes, the rGH gene is completely shut off in the hepatoma cells.