CAJ | 학술논문

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 Western blot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Western blot检测y-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)；Western blot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.
목적 관찰Ku80분자표체대간암세포DNA쌍련손상수평적영향.방법 Western blot검측40례간암화암주조직중γ-H2AX화Ku80표체수평；장Ku80기인전염도SMMC7721세포；면역형광화Western blot검측y-H2AX초점분포화단백표체.결과 γ-H2AX재간암조직중적표체여암주비교명현증고(31/40,77.5％),이Ku80표체명현감저(30/40,75％),γ-H2AX표체상조여Ku80표체하조상관(P＜0.05)；사선획득은정고표체Ku80적극륭세포；면역형광결과현시,Ku80 고표체극륭γ-H2AX초점수목여대조조비교명현감저(P＜0.01)；Western blot현시,Ku80고표체극륭γ-H2AX표체여대조조비교명현감저.결론 Ku80표체하조여간암세포적DNA쌍련손상상관,Ku80분자표체가감저간암세포DNA쌍련손상수평.
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 foci 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 cells or the vector-transfected cells.Western blotting further confirmed that the expression levels of γ-H2AX were decreased in the Ku80expressing cells compared with SMMC7721 or the vector-transfected cells.Conclusion Ku80 re-expression was able to decrease DNA double-strand breaks levels of hepatocellular carcinoma cells.