X射线对人外周血淋巴细胞DNA损伤修复和凋亡的影响 |
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引用本文: | 田铭,龚万军,徐向上,李小兰,陶德定,龚建平.X射线对人外周血淋巴细胞DNA损伤修复和凋亡的影响[J].国外医学:分子生物学分册,2010(6):508-512. |
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作者姓名: | 田铭 龚万军 徐向上 李小兰 陶德定 龚建平 |
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作者单位: | [1]华中科技大学同济医学院附属同济医院分子医学中心,胃肠外科武汉市430030 [2]广州军区武汉总医院心胸外科,武汉市430070 |
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基金项目: | 国家自然科学基金(No.30570908),国家重点基础研究发展规划项目(973计划)(No.2004CB518705) |
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摘 要: | 目的 研究不同时间诱导X射线照射的淋巴细胞进入细胞周期DNA损伤修复与凋亡的影响.方法 X射线(0.5 Gy)作用于正常人外周血淋巴细胞,以照射后不同时间点(0、4 h)分别加入PHA并分成两组,即照射后0 h加PHA组(A组)和照射后4 h加PHA组(B组),再分别培养0、0.5、2 h,用流式细胞术和免疫印迹法检测A组和B组γ-H2AX蛋白的表达,Annexin-V/PI法分析A、B两组的细胞凋亡率.结果 流式细胞术及免疫印迹结果均显示A组的γ-H2AX蛋白表达高于B组(P<0.05),且均先升高后降低.A组细胞凋亡率亦大于B组.结论 不同时间诱导被打击的淋巴细胞进入周期其可能发生DNA修复并同时伴随细胞凋亡的发生.
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关 键 词: | 电离辐射 植物凝集素 外周血淋巴细胞 γ-H2AX |
Influence of X-ray irradiation on DNA Damage Repair and Apoptosis in Peripheral Blood Lymphocytes |
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Authors: | TIAN Ming GONG Wanjun XU Xiangshang LI Xiaolan TAO Deding GONG Jianping |
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Institution: | 1 Molecular Medical Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, 430030, China;2 Department of Cardiothoracic Surgery, Wuha, General Hospital, Guangzhou Military Area Command, Wuhan, 430070, China) |
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Abstract: | Objective To investigate the effect of X-ray irradiation on DNA damage repair and apoptotic ratio in normal peripheral blood lymphocytes at difterent time points of addition of Phytohemagglatinin. Methods Phytohemagglutinin was added to lymphocytes at 0 h and 4 h after exposure to 0.5 Gy X- irradiation. Peripheral blood lymphocytes (PBLs) were divided into two groups, group A treated with PHA 0 h after irradiation and group B treated with PHA group 4 h after irradiation, and then incubated liar 0 h, 0.5 h and 2 h, respectively. Flow cytometry analysis and Western blotting were used to analyze the expression of γ-H2AX in the two groups. Apoptosis between the two groups was detected with annexinV-FITC/propidium iodide (PI) staining assay. Results Results of flow cytometry analysis and Western blot showed that 7-H2AX expression level in group A was generally higher than that in group B at either 0. 5 h or2 h time point (P〈0. 05), with a decrease at 2 h after irradiation. The apoptosis rate of group A was higher than that of group B. Conclusion Induction with phytohemagglutinin at different time points after X- irradiation of peripheral blood lymphocytes can repair the damaged lymphocytes, accompanied with oecurrence of apoptosis. |
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Keywords: | irradiation phytohemagglutinin peripheral blood lymphocyte γ-H2AX |
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