SIRT2沉默对肝癌细胞的迁移和侵袭能力的影响.docVIP

SIRT2沉默对肝癌细胞的迁移和侵袭能力的影响 宋春丽，任吉华，冉龙宽，李宛蔚，陈 娟 （400016重庆，重庆医科大学感染性疾病分子生物学教育部重点实验室） [摘要] 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（epithelial-mesenchymal 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。SIRT2基因沉默能抑制肝癌细胞的迁移和侵袭能力，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。 [关键词] SIRT2；肝癌；迁移；侵袭 [中图法分类号] [文献标志码] A Effect of SIRT2 on migration and invasion of hepatocellular carcinoma cells and its preliminary mechanism Song Chunli，Ren Jihua，Rang Longkuan，Li Wanyu，Chen Juan (Key Laboratory of Molecular Biology on Infection Diseases of Ministry of Education,Chongqing Medical University,Chongqing,400016,China) [Abstract]Objective To study the functional role of lentivirus-mediated silent information regulator 2(SIRT2) on migration and invasion of hepatocellular carcinoma cells (HCCs) and its preliminary mechanism． Methods The expression of SIRT2 in the HCC cell lines and L02 cells,an immortalized human live cell was analyzed by Real-time PCR and Western blot.SIRT2 expression was targeted by lentivirus-mediated shRNA interference technology and SIRT2 protein level was measured by Western blot．Effect of SIRT2 on migration and invasion of HCCs was detected by migration and invasion assay. EMT protein was measured by Western blot． Results SIRT2 was overexpressed in the HCC cell lines,Lentivirus-mediated shRNA significantly down-regulated the SIRT2 expression levels in HCCs．Knock down of SIRT2 suppressed the migration and invasion of HCCs . Knock down of SIRT2 in HCC cells induced the protein expression of epithelial markers E-cadherin and α-catenin that was accompanied by a concomitant re