八氯腺苷对A549和H1299细胞周期的影响及机制探讨  

Effect of 8-Chloro-adenosine on cell cycle of A549 and H1299 cells and its mechanism

作　　者：范芳[1] 安国顺[2] 李淑艳[2] 倪菊华[2] 贾弘禔[2] 

机构地区：[1]遵义医学院生物化学教研室,贵州遵义563099 [2]北京大学医学部生物化学与分子生物学系,北京100083

出　　处：《贵州医药》2014年第9期771-775,共5页Guizhou Medical Journal

基　　金：国家自然科学基金项目(No.30471975)

摘　　要：目的八氯腺苷对肺癌细胞株A549和H1299细胞增殖及细胞周期的影响及其机制探讨。方法 A549和H1299经10.0μmol/L八氯腺苷处理后,MTT法检测细胞生长和增殖情况,流式细胞术检测细胞周期,Western blot检测DNA-PKcs、γ-H2AX、Ku80、Ku70及TopoⅠ的表达情况。结果 MTT法检测结果显示,用10.0μmol/L八氯腺苷分别处理A549、H1299细胞24、48、72h后,A549、H1299细胞生长被抑制;流式细胞分析则显示,A549、H1299细胞G2/M期细胞数增多;Western blot结果表明,γ-H2AX的表达均增加,DNA-PKcs蛋白的表达均下降,而Ku80、Ku70的表达未见明显变化;经八氯腺苷处理后TopoⅠ蛋白在A549细胞中的表达逐渐增加,而在H1299细胞中的表达却逐渐下降。结论八氯腺苷可抑制A549和H1299细胞生长,诱导细胞发生G2/M期阻滞,其机制可能与DNA-PKcs表达下调有关。在A549细胞中TopoⅠ可能通过p53依赖途径介导DNA损伤修复,从而引发对细胞的毒性作用;而在p53缺失的H1299细胞中可能是通过其它途径调控细胞周期。Objective To explore the mechanisms of 8-chloro-adenosine-induced G2/M arrest in A549 and H1299 Cells .Methods The cell growth and cell cycle of A549 and H1299 were detected by MTT and flow cytometry analysis after 8-Cl-Ado exposures .The protein expression of DNA-PKcs ,γ-H2AX ,Ku80 ,Ku70 and Topo Ⅰ were detected by western blot （WB） assay .Results The MTT re-sults showed that the cell growth of A549 and H1299 cells were inhibited after 8-Cl-Ado exposures for 24 ,48 ,72 h .Flow cytometry analysis showed that the cells in G2/M phase of A549 and H1299 cells was increased .WB showed that the levels of γ-H2AX was upregulated ,DNA-PKcs was downregulat-ed in both A549 and H1299 cells ,meanwhile ,Ku80 and Ku70 expressions in the two cells did not see obvious change .The expression of Topo Ⅰ gradually increased in A549 cells ,and gradually decline in H1299 cells after 8-Cl-Ado treatment .Conclusion 8-Cl-Ado can inhibit the growth of A549 and H1299 cells ,induce cells in G2 /M phase retardation ,its mechanism may be related to DNA-PKcs expres-sion .Topo I in A549 cells may be mediated by p53 dependent way DNA damage repair ,causing toxic effect on cells .In loss of p53 H1299 cells may be initiate cell cycle arrest through other channels .

关 键 词：八氯腺苷 细胞周期 DNA依赖蛋白激酶催化亚基 拓扑异构酶I 

分 类 号：R734.2[医药卫生—肿瘤] R-33[医药卫生—临床医学]