吉林大学学报（医学版）2017，Vol.43Issue(3)：522-526,5.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
Effect of ionizing radiation on apoptosis of lung cancer H460 cells and its mechansim
摘要
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.关键词
ATRX/RNA干扰/caspase-3/辐射/肺肿瘤/细胞凋亡Key words
ATRX/RNAi/caspase-3/irradiation/lung neoplasm/apoptosis分类
医药卫生引用本文复制引用
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(3):522-526,5.基金项目
吉林省教育厅"十二五"科研规划项目资助课题(2014-192) （2014-192）
