博来霉素对肝癌细胞DNA损伤以及H2AX相互作用蛋白的影响  

The Effect of Bleomycin on DNA Damage and H2AX Interacting Proteins in Hepatocellular Carcinoma(HCC) Cells

作　　者：邹鹏[1,2] 杨晓丽[1,2] 陈先[1,2] 

机构地区：[1]复旦大学生物医学研究院,上海200032 [2]复旦大学化学系,上海200433

出　　处：《复旦学报（自然科学版）》2011年第2期172-177,共6页Journal of Fudan University：Natural Science

基　　金：国家高技术研究发展计划(863计划)资助项目(2006AA02A310);上海市科技创新团队计划资助项目(03DZ14024和07ZR14010)

摘　　要：组蛋白H2AX是DNA损伤修复反应中的关键蛋白,磷酸化的H2AX可以作为DNA损伤早期检测的金标准.实验通过检测磷酸化H2AX的改变来研究博来霉素对肝癌细胞和正常肝细胞DNA损伤作用的差异,推测肿瘤细胞抗药性可能是通过H2AX磷酸化来介导的.利用免疫共沉淀的方法,对博来霉素刺激下肝癌细胞中的H2AX复合物进行了免疫印迹的检验,结果显示DNA损伤修复相关蛋白PARP1和Ku70在DNA损伤后与H2AX相互作用出现了上调.14-3-3ζ是首次发现的H2AX相互作用蛋白,其与H2AX的相互作用在DNA损伤后也出现了增强.这些H2AX复合物在DNA损伤修复、细胞周期、细胞凋亡中扮演了重要角色,与肿瘤细胞对于放疗、化疗的效果有着密切的关系,构成了肿瘤细胞抗药性的分子基础,同时也为肿瘤的分子靶向治疗提供了可靠的依据.Histone H2AX plays a crucial role in DNA damage and repair response.Phosphorylated H2AX（γ-H2AX） was the gold standard for early detection of DNA damage.Here,the differences of Bleomycin（BLM）-induced DNA damage between hepatocellular carcinoma（HCC） cells and paired hepatocytes were investigated by measuring the amounts of γ-H2AX,implicating that the drug-resistant characteristic of HCC cells may be based on H2AX phosphorylation.Immunoprecipitation and Western blotting revealed that the interactions between H2AX and PARP1,or Ku70 were both increased under the stimulation of BLM in HCC cells.The interaction of 14-3-3ζ,a newly identified H2AX partners,with H2AX was also increased.These H2AX partners were involved in DNA damage and repair,cell cycle and apoptosis,and were also closely related to the effect of chemotherapy and radiotherapy in tumor cells,suggesting that these proteins may construct the molecular basis for drug-resistance of tumor cells,and provide the evidence of molecular target for tumor therapy.

关 键 词：博来霉素 肝癌 H2AX DNA损伤 

分 类 号：Q71[生物学—分子生物学]