异丙酚通过调节CIP2A/PP2A通路抑制HepG2细胞增殖

Propofol Inhibits the Proliferation of HepG2 Cells throughCIP2A/PP2A Pathway

目的探讨异丙酚抑制HepG2细胞增殖的作用及相关机制。方法将HepG2细胞分成未处理组、不同剂量(10、20、30μM)的异丙酚组;MTT法测定HepG2细胞的存活率;实时定量PCR检测调节蛋白磷酸酶2A的癌性抑制因子(cancerous inhibitor of protein phosphatase 2A,CIP2A)/蛋白磷酸酶2A(protein phosphatase 2A,PP2A)通路关键因子的基因表达;Wesrtern blot检测相关蛋白的表达水平;转染siRNA敲低CIP2A或过表达CIP2A后,给予不同浓度异丙酚,检测相关基因及蛋白表达水平。结果随着异丙酚浓度的增加,空载腺病毒(NC)组HepG2细胞增殖率受到抑制(P<0.05),当异丙酚浓度≥20μM时,CIP2A mRNA和蛋白表达量均低于NC组(P均<0.05);当异丙酚浓度≥10μM时,PP2A mRNA和蛋白的表达量均高于NC组(P均<0.05),c-Myc mRNA和蛋白的表达量低于NC组(P均<0.05)。在CIP2A敲低(Si-CIP2A)组,当异丙酚浓度≥10μM时,CIP2A mRNA和蛋白表达水平无变化(P均>0.05);而PP2A mRNA的表达水平升高(P<0.05),c-Myc mRNA表达量下降(P<0.05)。而CIP2A过表达(Ad-CIP2A)组,CIP2A、c-Myc和PP2A的mRNA在各异丙酚浓度组间差异均无统计学意义(P均>0.05)。结论异丙酚可以抑制肝癌细胞HepG2的增殖,可能与其调节CIP2A/PP2A通路有关。

Objective To explore the effect of propofol on the proliferation of HepG2 cells and related mechanisms.Methods HepG2 cells were divided into control group,three different concentrations of propofol groups,10μM,20μM,30μM respectively.MTT assay was performed to estimate the survival of HepG2 cells.Real-time PCR was performed to measure mRNA expression level of CIP2A/PP2A pathway related factors.Western blot was used to estimate related protein expression level.siRNA was transfected into HepG2 cells to knock down the expression of CIP2A,then related genes and proteins were measured.Results As the concentration of propofol increased,the proliferation rate of HepG2 cells in the NC group(no-load adenovirus group)was significantly inhibited(P<0.05).When the concentration of propofol was≥20μM,the expression of CIP2A mRNA and protein was significantly lower than NC group(P all<0.05),when the concentration of propofol≥10μM,the expression of PP2A mRNA and protein was significantly higher than that of NC group(P all<0.05),and the expression of c-Myc mRNA and protein was significantly lower than that of NC group(P all<0.05).After siRNA knocked down CIP2A,in the Si-CIP2A group(CIP2A knockdown group),when the concentration of propofol was≥10μM,the expression levels of CIP2A mRNA and protein changed significantly(P all>0.05);while the expression levels of PP2A mRNA increased significantly(P<0.05),c-Myc mRNA expression decreased significantly(P<0.05).However,in the Ad-CIP2A group(CIP2A overexpression group)after CIP2A overexpression,the mRNAs of CIP2A,c-Myc and PP2A were not statistically different among the propofol concentration groups(P all>0.05).Conclusion Propofol can inhibit the proliferation and growth of hepato cellular carcinoma cells via regulating CIP2A/PP2A pathway.