沉默信息调节因子3对肝癌细胞增殖的影响  

Role of SIRT3 in regulating proliferation of hepatocellular carcinoma cells in vitro

作　　者：陶颖[1] 陈娟[2] 

机构地区：[1]中国人民解放军第161医院,湖北武汉430012 [2]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《南方医科大学学报》2016年第2期195-199,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81472271;81201282)~~

摘　　要：目的探讨沉默信息调节因子3（SIRT3）对肝癌细胞增殖的影响。方法 Western blotting检测SIRT3在正常肝组织、永生化肝细胞及肝癌细胞系中的蛋白表达水平;在肝癌细胞中过表达SIRT3,台盼蓝排斥实验检测过表达SIRT3对肝癌细胞增殖的影响,Ed U标记实验检测SIRT3对肝癌细胞DNA合成的影响;在肝癌细胞中过表达SIRT3,平板集落实验检测SIRT3对肝癌细胞集落形成能力的影响;q RT-PCR验证SIRT3的沉默效率,台盼蓝排斥实验检测SIRT3沉默对肝癌细胞增殖的影响,Ed U标记实验检测SIRT3沉默对肝癌细胞DNA合成的影响。结果 SIRT3在肝癌细胞系中的蛋白水平较永生化肝细胞、正常肝组织降低;SIRT3在肝癌细胞中成功过表达,过表达SIRT3使肝癌细胞增殖数目减少了约为51%~61%（P〈0.001）,使肝癌细胞DNA合成下降了约57%（P〈0.05）;过表达SIRT3抑制肝癌细胞的集落形成能力;SIRT3沉默有效,SIRT3沉默使肝癌细胞的增殖增加了约51%~61%（P〈0.01）,使肝癌细胞的DNA合成能力增强了137%~149%（P〈0.01）。结论 SIRT3可抑制肝癌细胞的增殖。Objective To explore the role of SIRT3 in regulating the proliferation of hepatocellular carcinoma（HCC） cells in vitro.Methods The protein expression of SIRT3 in 2 normal liver tissues, 2 immortalized hepatocyte lines, and 3 HCC cell lines was determined with Western blotting. SIRT3 overexpression and knockdown in HCC cells were induced by transfection with a vector expressing SIRT3 and a si RNA construct targeting SIRT3, respectively. The efficiency of SIRT3 overexpression and knockdown was detected by Western blot and q RT- PCR, respectively. The proliferation of the transfected HCC cells was examined using Trypan blue exclusion assay, and the cellular DNA synthesis was tested using Ed U incorporation assay. The colony-forming ability of the cells was analyzed by colony formation assays. Results SIRT3 expression was significantly lower in the 3 HCC cell lines than in immortalized hepatocytes and normal liver tissues. SIRT3 overexpression in HCC cells significantly lowered the cell proliferation by 51%- 61%（P〈0.001）, reduced cellular DNA synthesis by 57%（P〈0.05）, and inhibited colony formation of the cells. SIRT3 knockdown significantly increased the proliferation of HCC cells by 51%-61%（P〈0.01） and enhanced DNA synthesis by 137%- 149%（P〈0.01）. Conclusion SIRT3 plays a inhibitory role in regulating the proliferation of HCC cells in vitro.

关 键 词：沉默信息调节因子3 肝癌 增殖 

分 类 号：R735.7[医药卫生—肿瘤]