REGULATION OF LOW-DENSITY-LIPOPROTEIN RECEPTORS IN THE HUMAN HEPATOMA-CELL LINE HEP-G2 - EFFECT OF PHORBOL 12-MYRISTATE 13-ACETATE AND LOW-DENSITY-LIPOPROTEIN

The regulation of low-density-lipoprotein(LDL)-receptor activity by 4beta-phorbol 12-myristate 13-acetate (PMA) and LDL was investigated in the human hepatoma cell line Hep G2. Treatment of Hep G2 cells for 22 h with PMA results in an 18.6-fold increase in the amount of LDL-binding sites on the cell surface. The rate of turnover of LDL receptors was not significantly altered upon PMA treatment. Treatment of cultured rat parenchymal cells and human parenchymal cells with PMA did not lead to increased binding of LDL to these cells, suggesting that protein-kinase-C-mediated regulation of LDL-receptor activity is specific for Hep G2 cells and that in this aspect of regulation of LDL receptors, Hep G2 cells do not reflect human hepatocytes. The down-regulation of LDL receptors by a 22-h prior incubation with LDL in PMA-treated Hep G2 cells, in which LDL receptors are upregulated, is more effective than in non-treated cells. Prior incubation of control Hep G2 cells with an excess of LDL caused a partial down-regulation to 33% of the initial level of receptor binding. In PMA-treated Hep G2 cells an excess of non-labeled LDL, led to a down-regulation to 13% of the PMA-induced level. Prior incubation of Hep G2 cells with LDL in the presence of PMA led to a 2.3-fold increase of intracellular cholesteryl esters and a 9.1-fold increase in acyl-CoA:cholesterol-acyltransferase (ACAT) activity. In control cells, LDL prior incubation led to a 1.6-fold increase in intracellular cholesteryl esters and a 1.8-fold increase of ACAT activity. It is concluded that in Hep G2 cells LDL itself can be an effective suppressor of the expression of LDL receptors, provided that the initial amount of receptor allows an adequate intracellular delivery of cholesterol to its sterol-regulatory site.