电离辐射对肺癌H460细胞凋亡的影响及其机制

发布时间：2018-06-01 18:12

  本文选题：ATRX + RNA干扰　； 参考：《吉林大学学报(医学版)》2017年03期

【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shATRX3-H460组,分别给予0、2和8Gy 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8Gy X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(P0.05或P0.01);8Gy X线照射后sh-ATRX3-H460组细胞凋亡率明显高于sh-control-H460组(P0.01);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,8Gy 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。
[Abstract]:Aim: to study the effect of ionizing radiation on apoptosis of H460 cells and its mechanism by silencing the ATRX gene of H460 cells by RNA interference RNAi. Methods: the lentivirus expression vector targeting ATRX gene was transfected into 293T cells. The cell model of ATRX silencing was obtained by puromycin positive screening. The cells were named sh-ATRX1-H460h-ATRX2-H460 and sh-ATRX3-H460 cells, and sh-control-H460 cells were used as control. According to the silencing results, they were divided into sh-control-H460 group and shATRX3-H460 group. ATRX, polyadenylate ribonucleotidyl polymerase 1 (PARP1), caspase-3 protein expression and Annexin 鈪，

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