Progesterone stimulation of HMG-CoA reductase activity in cultured cells

In a previous study we showed that progesterone (PG) stimulated HMG-CoA reductase (HMGR) activity in rat intestinal epithelial cells (IEC-6) incubated in the presence or absence of low-density lipoprotein (LDL) [1,2]. In the present study we examined further the mechanism of this stimulation. We observed that the stimulation of HMGR activity by PG was completely prevented by cycloheximide. Turnover studies utilizing immunoprecipitation of HMGR-labeled with [35S]methionine revealed that PG increased reductase activity by inhibiting HMGR degradation without affecting the synthesis of HMGR. The stimulation of HMGR activity by progesterone could be accounted for by a continuous synthesis of HMGR while its degradation was retarded. In the presence of LDL, the activity of HMGR in IEC-6 cells was effectively inhibited, however PG was able to stimulate HMGR in the presence of LDL. This effect was not due to an interference of normal cellular metabolism of LDL, since PG had no effect on the cellular uptake and lysosomal degradation of 125I-LDL. PG did not affect of the rate of lysosomal hydrolysis of [3H]cholesteryl linoleate-LDL. The free [3H]cholesterol derived from [3H]cholesteryl linoleate-LDL moved to the cell membrane and effluxed to HDL3 in the medium at the same rate in the presence or absence of PG. Although PG did not affect LDL metabolism, pre-treatment of cells with LDL delayed the onset of HMGR stimulation by PG. In IEC-6 cells deprived of LDL for 24 h, the HMGR activity was stimulated immediately following PG addition. In cells pre-treated with LDL for 24 h, the stimulation was delayed by 4 h. Treatment of cells with 25-hydroxycholesterol completely prevented PG stimulation of HMGR activity. We propose that the stimulation of HMGR activity in the presence or absence of LDL is related to the ability of PG to attenuate the formation and/or action of intracellular HMGR repressor molecules which accelerate the degradation of HMGR.