PIK3R3 Regulates Lipid Metabolism and Proliferation of Hepatocellular Carcinoma Cells by Regulating PPARA / 华中科技大学学报(医学版)

ABSTRACT

Objective@#To explore whether PIK3R3 is dependent on PPARA to regulate fat metabolism and proliferation of liver cancer cells.@* Methods@#The PIK3R3 overexpression plasmids and its specific siRNA were transfected into hepatocellular carcinoma cells HepG2 and SMMC-7721. The level of PPARA was detected by Western blotting（WB）and the ketone body content in supernatant was measured by colorimetric method.DNA synthesis and proliferation of hepatocellular carcinoma cells were detected by BrdU/PI incorporation method and CCK8 method. Then we used PIK3R3 overexpression or silenced hepatocellular carcinoma cell lines to observe the effect of PIK3R3 on the proliferation of hepatocellular carcinoma cells by plate cloning experiments. We intervened the expression of PIK3R3 and then observed the lipid metabolism and cell proliferation，so as to determine whether the role of PIK3R3 in hepatocellular carcinoma cells depends on PPARA. Finally, we added MG132（proteasome inhibitor）or CHX（cycloheximide）to PIK3R3 overexpression or silencing hepatocellular carcinoma cells to detect the level of PPARA by WB，next we constructed the plasmids of PPARA promoter luciferases and observed the effect of PIK3R3 on the activity of PPARA promoter. @*Results@#PIK3R3 influenced the expression of PPARA，lipid metabolism and proliferation of hepatocellular carcinoma cells，and PPARA partly reversed the effects of PIK3R3.@*Conclusion@#PIK3R3 can regulate the fat metabolism and proliferation of hepatoma cells by regulating the expression of PPARA.