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该文旨在探讨沉默信息调节因子3(sirtuin 3,SIRT3)对肝癌细胞凋亡的影响,并研究SIRT3调节肝癌细胞凋亡的分子机制。运用流式细胞术检测SIRT3过表达对肝癌细胞系(SMMC-7721和SK-Hep-1)凋亡的影响;通过si RNA靶向沉默SIRT3并检测SIRT3沉默对肝癌细胞凋亡的影响;实时荧光定量PCR(Real-time PCR)分析SIRT3对Bcl-2家族成员m RNA水平的影响,筛选受SIRT3调节的Bcl-2家族成员;Western blot进一步检测SIRT3对目标Bcl-2家族成员蛋白水平的影响;流式细胞术分析目标Bcl-2家族成员在SIRT3诱导肝癌细胞凋亡中的作用。结果显示,SIRT3过表达促进肝癌细胞凋亡并引起Bax mRNA和蛋白水平升高;SIRT3沉默抑制肝癌细胞凋亡,同时也抑制Bax蛋白水平表达,Bax沉默显著减少了SIRT3过表达细胞中的凋亡数目。该研究结果提示,SIRT3通过凋亡调节基因Bax诱导肝癌细胞凋亡。
The aim of this study was to investigate the effect of Sirtuin 3 (SIRT3) on hepatocellular carcinoma cell apoptosis and to investigate the molecular mechanism by which SIRT3 regulates hepatoma cell apoptosis. The effect of SIRT3 overexpression on the apoptosis of hepatocellular carcinoma cell lines (SMMC-7721 and SK-Hep-1) was detected by flow cytometry. SIRT3 was silenced by si RNA and the effect of SIRT3 silencing on the apoptosis of hepatoma cells was detected. Real-time fluorescence The effect of SIRT3 on the m RNA level of Bcl-2 family members was analyzed by Real-time PCR and the Bcl-2 family members regulated by SIRT3 were screened. The effect of SIRT3 on the protein levels of target Bcl-2 family members was further detected by Western blot. Flow Cytometry Analysis of the Role of Target Bcl-2 Family Members in SIRT3-Induced Apoptosis of Hepatoma Cells. The results showed that SIRT3 overexpression promoted hepatocarcinoma cell apoptosis and increased Bax mRNA and protein levels. SIRT3 silencing inhibited hepatocellular carcinoma cell apoptosis as well as Bax protein expression, and Bax silencing significantly reduced apoptosis in SIRT3 overexpressing cells number. The results suggest that SIRT3 induces hepatocellular carcinoma cell apoptosis through the apoptosis-regulating gene Bax.