RENJing-hua,LIN Ju-sheng,LIU Yao.γ-H2AX assay as a measurement of DNA double-strand breaks in irradiated HepG2 cells[J].Chinese Journal of Radiological Medicine and Protection,2007,27(5):450-454
γ-H2AX assay as a measurement of DNA double-strand breaks in irradiated HepG2 cells
Received:October 19, 2006  
DOI:
KeyWords:γ-H2AX  DNA double-strand breaks  Ionizing radiation
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Author NameAffiliationE-mail
RENJing-hua Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China  
LIN Ju-sheng Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China jslin@tjh.tjmu.edu.cn 
LIU Yao Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China  
陈琼 Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China  
董旭 Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China  
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Abstract::
      Objective To explore the possibility of detecting DNA double-strand breaks (DSBs) in irradiated HepG2 cells by γ-H2AX assay and to investigate the γ-H2AX expression in different cell cycle phases. Methods HepG2 cells were blocked by TdR in G1 phase of cell cycle,and then cultured for different hours after TdR was removed. Synchronous S phase cells and G2/M phase cells were collected respectively and irradiated with 60Co γrays. Western blotting and immunofluorescence were used to analyze the expression ofγ-H2AX in cells. Results Synchronous S phase cells and G2/M phase cells were obtained at hours 34 and 40 post TdR block respectively. γ-H2AX level increased rapidly after cells were exposed to 60Co γ rays, especially in S phase of the cell cycle. Conclusions The cells in S phase are more prone to be damaged by ionizing radiation than those in G2/M.γ-H2AX assay can be regarded as an early diagnosis indicator of DSBs.
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