依匹哌唑对结肠癌细胞系HCT116和SW620细胞内胆固醇合成的影响及其机制

Effect and mechanism of brexpiprazole on cholesterol synthesis of human colorectal cancer cell lines HCT116 and SW620

目的观察依匹哌唑对结肠癌细胞系HCT116和SW620细胞内胆固醇合成的影响,并探讨其机制。方法取对数生长期HCT116、SW620细胞,分别分为依匹哌唑组和对照组,依匹哌唑组加入含20μmol/L依匹哌唑的培养基,对照组加入等量完全培养基。两组细胞继续培养24 h后,采用微板法检测细胞中总胆固酮(TC),采用实时荧光定量PCR法检测人3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)m RNA、人3-羟基-3-甲基戊二酰辅酶A合成酶1(HMGCS1)m RNA,采用Western Blotting法检测HMGCR、HMGCS1、p-PI3K、p-AKT、SREBP2蛋白。结果依匹哌唑组HCT116、SW620细胞中TC含量均低于对照组(P均<0.05)。依匹哌唑组HCT116、SW620细胞中HMGCR m RNA、HMGCS1 m RNA相对表达量均低于对照组(P均<0.05)。依匹哌唑组HCT116、SW620细胞中HMGCR、HMGCS1、p-PI3K、p-AKT、SREBP2蛋白相对表达量均低于对照组(P均<0.05)。结论依匹哌唑可抑制HCT116和SW620细胞的细胞内胆固醇合成,其机制可能与依匹哌唑抑制PI3K/Akt-SREBP2信号通路蛋白的表达有关。

Objective To observe the effect of brexpiprazole on cholesterol synthesis in colorectal cancer cell lines HCT116 and SW620,and to explore its mechanism.Methods HCT116 and SW620 cells in the logarithmic growth phase were divided into the brexpiprazole group and control group,respectively.Cells in the brexpiprazole group were add⁃ed with medium containing 20μmol/L brexpiprazole,while cells in the control group were added with equal amount of me⁃dium.After the cells were cultured for 24 h,the total cholesterol(TC)in the cells was detected by microplate assay.Hu⁃man 3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR)mRNA and human 3-hydroxy-3-methylglutaryl-CoA synthase 1(HMGCS1)mRNA were detected by real-time fluorescence quantitative PCR.HMGCR,HMGCS1,p-PI3K,p-AKT,and SREBP2 proteins were detected by Western blotting.Results The TC levels of HCT116 and SW620 cells were lower in the brexpiprazole group than in the control group(both P<0.05).The relative expression levels of HMGCR mRNA and HMGCS1 mRNA in HCT116 and SW620 cells in the brexpiprazole group were lower than those in the control group(all P<0.05).The relative protein expression levels of HMGCR,HMGCS1,p-PI3K,p-AKT and SREBP2 in HCT116 and SW620 cells of the brexpiprazole group were lower than those of the control group(all P<0.05).Conclusion Brexpipra⁃zole inhibits intracellular cholesterol synthesis in HCT116 and SW620 cells possibly by inhibiting the protein expression of PI3K/Akt-SREBP2 signaling pathway.