电离辐射对肺癌H460细胞凋亡的影响及其机制

doi:10.13481/j.1671-587x.20170312

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (03): 522-526.doi: 10.13481/j.1671-587x.20170312

电离辐射对肺癌H460细胞凋亡的影响及其机制

张晶¹, 王志成^2,3, 赵大力⁴, 卢晓倩², 沈智渊³, 齐亚莉^1,2

1. 北华大学公共卫生学院流行病学教研室, 吉林吉林 132011;
2. 吉林大学公共卫生学院卫生部放射生物学重点实验室, 吉林长春 130021;
3. 美国罗格斯新泽西州立大学肿瘤研究所, 新泽西州皮斯卡塔韦 08903-2681;
4. 吉林省出入境检验检疫局, 吉林长春 130013

收稿日期:2016-09-02 出版日期:2017-05-28 发布日期:2017-06-01
通讯作者: 王志成,副教授,硕士研究生导师(Tel:0431-85619443,E-mail:zhicheng@jlu.edu.cn);齐亚莉,副教授,硕士研究生导师(Tel:0431-85619443,E-mail:374494617@qq.com) E-mail:zhicheng@jlu.edu.cn;374494617@qq.com
作者简介:张晶(1973-),女,吉林省长春市人,副教授,医学硕士,主要从事肿瘤流行病学方面的研究。
基金资助:
吉林省教育厅"十二五"科研规划项目资助课题（2014-192）

Effect of ionizing radiation on apoptosis of lung cancer H460 cells and its mechansim

ZHANG Jing¹, WANG Zhicheng^2,3, ZHAO Dali⁴, LU Xiaoqian², SHEN Zhiyuan³, QI Yali^1,2

1. Department of Epidemiology, School of Public Health, Beihua University, Jilin 132011, China;
2. Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China;
3. Cancer Institute, State University of New Jersey, New Jersey 08903-2681, USA;
4. Jilin Province Entry and Exit Inspection and Quarantine Bureau, Changchun 130013, China

Received:2016-09-02 Online:2017-05-28 Published:2017-06-01

摘要/Abstract

摘要： 目的：以RNA干扰（RNAi）沉默肺癌H460细胞ATRX基因，探讨电离辐射对肺癌H460细胞凋亡的影响及其作用机制。方法：以靶向ATRX基因的慢病毒表达载体转染293T细胞，所得慢病毒2次感染肺癌细胞H460，通过puromycin阳性筛选获得ATRX 沉默的细胞模型，命名为sh-ATRX1-H460、sh-ATRX2-H460和sh-ATRX3-H460细胞，以sh-control-H460细胞作为对照。根据沉默结果分为sh-control-H460组和sh-ATRX3-H460组，分别给予0、2和8 Gy X射线照射。Western blotting法检测细胞中ATRX、多聚腺苷酸二磷酸核糖基聚合酶1（PARP1）和caspase-3蛋白表达量；AnnexinⅤ-FITC/PI试剂盒和流式细胞术检测各组细胞凋亡率。结果：成功获得ATRX沉默的肺癌sh-ATRX3- H460细胞模型。与照射前比较，2和8 GyX射线照射后sh-control-H460组细胞中ATRX蛋白表达量增加，sh-ATRX3-H460组细胞中无ATRX蛋白表达；与照射前比较，照射后sh-control-H460组和sh-ATRX3-H460组细胞凋亡率均明显升高（P<0.05或P<0.01）；8 Gy X线照射后sh-ATRX3- H460组细胞凋亡率明显高于sh-control-H460组（P<0.01）；2组细胞中cleaved PARP1蛋白表达量均增加且规律相似；sh-control-H460组细胞中procaspase-3蛋白表达量无明显变化，8 Gy X射线照射后sh-ATRX3-H460组细胞中procaspase-3蛋白表达量明显增加。结论：RNAi可以实现ATRX沉默，ATRX沉默能够增加辐射诱导的细胞凋亡，其机制可能与PARP1-caspase-3通路有关联。

关键词: caspase-3, ATRX, 辐射, 肺肿瘤, 细胞凋亡, RNA干扰

Abstract: Objective: To explore the effect of ionizing radiation on apoptosis of lung cancer H460 cells after ATRX was silenced by RNAi and its mechanism. Methods: The lentivirus expression vectors targeting ATRX were transfected into the 293T cells,and the lung cancer H460 cells were infected with lentivirus twice,and the ATRX silenced cell model was obtained after puromycin positive screening,then they were named as sh-ATRX1-H460,sh-ATRX2-H460,and sh-ATRX3-H460 cells; the sh-control-H460 cells were regarded as control cells.The cells were divided into sh-control-H460 group and sh-ATRX3-H460 group,accroding to the silencing results and were irradiated by 0,2 and 8 Gy X-rays.The expression levels of ATRX,poly(ADP-ribose) polymerase 1(PARP1),and caspase-3 proteins were measured by Western blotting method; the apoptotic rate was measured by flow cytometry and AnnexinⅤ-FITC/PI kits. Results: The lung cancer cell model of sh-ATRX3-H460 silenced by ATRX was obtained successfully.After 2 and 8 Gy X-ray irradiation,compared with before irradiation, the expression level of ATRX protein in sh-control-H460 group was increased,while there was no expression of ATRX protein in sh-control-H460 group;compared with before irradiation, the apoptotic rates of cells in two groups were increased(P<0.05 or P<0.01);the apoptotic rate in sh-ATRX3-H460 group was significantly higher than that in sh-control-H460 group after 8 Gy X-ray irradiation (P<0.01).The expressions of cleaved PARP1 in the cells in both two groups after 2 Gy and 8 Gy X-ray irradiation were increased and showed similar rule.The expression level of procaspase-3 protein in sh-control-H460 group had little change,and it was increased significantly in sh-ATRX3-H460 group after 8 Gy X-ray irradiation. Conclusion: ATRX silencing can be achived by RNAi,then the silencing could increase the apoptosis induced by irradiation and its mechanism may be related to the PARP1-caspase-3 pathway.

Key words: caspase-3, ATRX, irradiation, lung neoplasm, apoptosis, RNAi

中图分类号:

R144.1

张晶, 王志成, 赵大力, 卢晓倩, 沈智渊, 齐亚莉. 电离辐射对肺癌H460细胞凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2017, 43(03): 522-526.

ZHANG Jing, WANG Zhicheng, ZHAO Dali, LU Xiaoqian, SHEN Zhiyuan, QI Yali. Effect of ionizing radiation on apoptosis of lung cancer H460 cells and its mechansim[J]. Journal of Jilin University Medicine Edition, 2017, 43(03): 522-526.

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电离辐射对肺癌H460细胞凋亡的影响及其机制

Effect of ionizing radiation on apoptosis of lung cancer H460 cells and its mechansim

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