Ku80分子表达对肝癌细胞DNA双链损伤水平的影响  

The effect of Ku80 re-expression on the DNA double-strand breaks in hepatocellular carcinoma cells

作　　者：魏双[1] 黄志勇[1] 刘杨安[1] 王洋洋[1] 纪桂宝[1] 占大钱[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科中心,武汉430030

出　　处：《中华实验外科杂志》2011年第10期1646-1648,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30772126、30972901）;教育部新世纪人才支持计划资助项目（NCET-04-0701）

摘　　要：目的观察Ku80分子表达对肝癌细胞DNA双链损伤水平的影响。方法Westernblot检测40例肝癌和癌周组织中γ-H2AX和Ku80表达水平；将Ku80基因转染到SMMC7721细胞；免疫荧光和Westernblot检测γ-H2AX焦点分布和蛋白表达。结果γ-H2AX在肝癌组织中的表达与癌周比较明显增高（31／40，77．5％），而Ku80表达明显减低（30／40，75％），γ-H2AX表达上调与Ku80表达下调相关（P〈0．05）；筛选获得稳定高表达Ku80的克隆细胞；免疫荧光结果显示，Ku80高表达克隆γ-H2AX焦点数目与对照组比较明显减低（P〈0．01）；Westernblot显示，Ku80高表达克隆γ-H2AX表达与对照组比较明显减低。结论Ku80表达下调与肝癌细胞的DNA双链损伤相关，Ku80分子表达可减低肝癌细胞DNA双链损伤水平。Objective To investigate the effect of Ku80 re-expression on DNA double-strand breaks in HCC cells. Methods γ-H2AX and Ku80 expressive levels in HCC tissues and corresponding liver tissues were analyzed by the Western blotting. PcDNA3.1 （ ＋ ） -myc-his-Ku80 and pcDNA3.1 （ ＋ ） - myc-his expressive plasmids were transfected into Ku80 deficient SMMC7721 to generate Ku80-expressing SMMC7721 stable clones through G418 screen. γ-H2AX assay was used to quantify the numbers of γ-H2AX loci in the nuclei of Ku80-expressing clones and control cells, while Western blotting was used to compare the γ-H2AX expressive levels in Ku80-expressing clones and control cells. Results The γ-H2AX expressions in HCC tissues were higher than those in the corresponding adjacent liver tissues （31/40, 77.5% ）, while Ku80 expressions were found to be decreased or lost in HCC tissues compared with adjacent liver tissues （30/40, 75% ） ; Significant correlation was found between the decreased expression of Ku80 and increased γ-H2AX in HCC（P 〈0. 05）. The Western blotting analysis confirmed that Ku80-transfected clones expressed high protein levels of Ku80, whereas vector-transfected clone and the parental SMMC7721 cells lacked Ku80 expression. γ-H2AX assay indicated that the numbers of γ-H2AX foci in the nuclei of Ku80-expressing clone cells were significantly decreased compared with SMMC7721 ceils or the vector-transfected cells. Western blotting further confirmed that the expression levels of γ-H2AX were decreased in the Ku80- expressing cells compared with SMMC7721 or the vector-transfected cells. Condusion Ku80 re-expression was able to decrease DNA double-strand breaks levels of hepatocellular carcinoma cells.

关 键 词：KU80 人肝细胞癌 Γ-H2AX DNA双链损伤 

分 类 号：R735.7[医药卫生—肿瘤]