Sirtuin 6对肝癌细胞增殖的影响  

Effect of sirtuin 6 on proliferation of hepatocellular carcinoma cells

作　　者：陶娜娜[1] 周洪钟[1] 任吉华[1] 陈祥[1] 李宛蔚[1] 刘波[1] 陈娟[1] 

机构地区：[1]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：《中国病理生理杂志》2016年第6期1031-1036,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81472271)

摘　　要：目的:探究sirtuin 6(SIRT6)对肝癌细胞增殖的影响。方法:RT-q PCR检测200例肝癌病人及50例健康人外周血中的SIRT6的mRNA表达水平,并将肝癌病人外周血SIRT6的mRNA表达水平与多个临床病理参数相结合进行统计学分析。Western blotting检测SIRT6在原代肝细胞、永生化肝细胞及肝癌细胞系中的蛋白表达水平。在肝癌细胞中利用shRNA干扰SIRT6基因的表达,并通过Western blotting验证沉默效率;MTS实验检测SIRT6基因沉默对肝癌细胞活力的影响。Ed U标记实验检测SIRT6基因沉默对肝癌细胞DNA合成的影响;平板集落实验检测SIRT6基因沉默对肝癌细胞集落形成能力的影响;软琼脂集落形成实验检测SIRT6基因沉默对肝癌细胞锚定非依赖生长能力的影响。结果:SIRT6在肝癌病人的外周血中表达明显增高,且SIRT6的表达量增高与肿瘤的大小、肿瘤的分级以及肿瘤的血管侵袭相关。进一步验证发现SIRT6在肝癌细胞系中表达水平较原代肝细胞PHH及永生化肝细胞MIHA增高。在2个肝癌细胞系中沉默SIRT6可以抑制肝癌细胞的活力及DNA合成能力,也可抑制肝癌细胞集落形成能力及锚定非依赖生长能力。结论:SIRT6促进肝癌细胞的增殖及恶性转化。AIM： To illuminate the effect of sirtuin 6（ SIRT6） on the proliferation of hepatocellular carcinoma（ HCC） cells. METHODS： The mRNA expression of SIRT6 in the peripheral blood from 200 cases of HCC patients and50 cases of healthy people was detected by real-time quantitative PCR（ RT-q PCR）. The mRNA expression levels of SIRT6 in the peripheral blood from 200 cases of HCC patients were combined with multiple clinicopathologic parameters for statistical analysis. The protein expression of SIRT6 in primary hepatocytes,immortalized hepatocytes and 4 hepatoma cell lines were determined by Western blotting. The silencing of SIRT6 was conducted by transfection of vector expressing short hairpin RNA targeting on SIRT6,and the protein level of SIRT6 was measured by Western blotting. The viability of HCC cells was tested by MTS assay. DNA synthesis was analyzed by Cell-LightTMEd U Apollo 488 In Vitro Imaging Kit. The abilities of colony formation and anchorage-dependent growth were measured by colony formation assay and soft agar assay,respectively. RESULTS： The mRNA expression of SIRT6 in the peripheral blood of HCC patients was significantly higher than that in the healthy people,and its expression was highly associated with tumor size,tumor grade and vascular invasion.SIRT6 expression in 4 hepatoma cell lines was significantly higher than that in the others. SIRT6 silencing led to a significant decrease in the cell viability as tested by MTS assay. Ed U staining revealed that SIRT6 silencing reduced DNA synthesis. SIRT6 silencing reduced the ability of colony formation and anchorage-dependent growth as determined by colony formation assay and soft agar assay,respectively. CONCLUSION： Sirtuin 6 promotes the proliferation and malignant transformation of HCC cells.

关 键 词：SIRTUIN 6 肝细胞癌 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤] R730.23[医药卫生—临床医学]