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目的探讨慢病毒介导的沉默信息调节因子2(silent information regulator2,SIRT2)基因沉默对肝癌细胞的迁移和侵袭能力的影响。方法 Western blot分析SIRT2在多个肝癌细胞系和永生化肝细胞中的表达,通过慢病毒介导的shRNA干扰技术靶向沉默SIRT2的表达,并通过Real-time PCR和Western blot验证SIRT2基因的沉默效果。细胞迁移及侵袭实验检测SIRT2基因沉默对肝癌细胞迁移和侵袭能力的影响,Western blot检测上皮细胞间充质转化(epithelialmesenchymal transition,EMT)指标蛋白(E-cadherin、α-catenin、α-SMA、N-cadherin)的表达。结果 SIRT2在多个肝癌细胞系中蛋白表达水平明显上调;慢病毒介导的shRNA能显著抑制细胞SIRT2的表达水平(P<0.05)。SIRT2基因沉默能抑制肝癌细胞的迁移和侵袭能力(P<0.01),并使上皮属性蛋白E-cadherin、α-catenin表达水平升高,使间质属性蛋白α-SMA、N-cadherin表达水平降低。结论 SIRT2基因沉默可以通过抑制肝癌细胞上皮细胞间充质转化从而抑制肝癌细胞的迁移和侵袭能力。
Objective To investigate the effect of lentivirus-mediated silent information regulator2 (SIRT2) gene silencing on the migration and invasion of hepatocellular carcinoma cells. Methods The expression of SIRT2 in multiple hepatocellular carcinoma cell lines and immortalized hepatocytes was analyzed by Western blot. The silencing SIRT2 expression was targeted by lentivirus-mediated shRNA interference. SIRT2 gene silencing was confirmed by Real-time PCR and Western blot effect. The effects of SIRT2 gene silencing on migration and invasion ability of hepatocellular carcinoma cells were detected by cell migration and invasion assay. Western blot was used to detect the expression of E-cadherin (α-catenin, α-SMA, N -cadherin) expression. Results The protein expression of SIRT2 in multiple hepatocellular carcinoma cell lines was significantly up-regulated. The lentivirus-mediated shRNA significantly inhibited the expression of SIRT2 in cells (P <0.05). SIRT2 silencing could inhibit the migration and invasion of hepatocellular carcinoma cells (P <0.01), and increase the expression of epithelial protein E-cadherin and α-catenin, and decrease the expression of interstitial protein α-SMA and N-cadherin . Conclusion SIRT2 gene silencing can inhibit the migration and invasion ability of hepatocellular carcinoma cells by inhibiting the mesenchymal transition of hepatoma cells.