pEgr1-hsTRAIL质粒对肺腺癌A549细胞放射敏感性及DR4和DR5表达水平的影响  

Influence of pEgr1-hsTRAL plasmid on radiosensitivity and DR4 and DR5 expression levels in lung adenocarcinoma A549 cells

作　　者：杨艳明[1] 方芳[2] 刘念[1] 曲志杰[1] 王志成[2] 张艳秋[3] 

机构地区：[1]吉林大学第二医院肿瘤生物治疗中心,吉林长春130041 [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]吉林大学第二医院手术室,吉林长春130041

出　　处：《吉林大学学报（医学版）》2013年第2期199-203,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30901702);吉林大学科学前沿与交叉学科创新项目资助课题(201103052)

摘　　要：目的:检测转染pEgr1-hsTRAIL质粒的人肺腺癌A549细胞放射敏感性变化及其对死亡受体(DR)4和DR 5mRNA和蛋白表达的影响,探讨pEgr1-hsTRAIL质粒的辐射增敏效应和诱导细胞凋亡的可能机制。方法:实验分为正常对照(Normal control)组、pEgr1-hsTRAIL组、6Gy X线照射组和pEgr1-hsTRAIL+6GyX线照射组。A549细胞经过阳离子脂质体转染、X线深部治疗机照射后,利用平板克隆形成试验检测细胞放射敏感性,反转录RCR(RT-PCR)法检测DR4和DR5mRNA表达变化,Western blotting法检测DR4和DR5蛋白表达的变化。结果:正常对照组和pEgr1-hsTRAIL组A549细胞的平均致死剂量(D0值)分别为3.26和1.91Gy,说明pEgr1-hsTRAIL质粒能够增强A549细胞的放射敏感性。RT-PCR显示,转染pEgr1-hsTRAIL质粒对DR4和DR5mRNA和蛋白表达水平无明显影响,而6Gy X线照射后DR4和DR5mRNA表达均显著高于正常对照组(P<0.05),转染pEgr1-hsTRAIL质粒后再进行6Gy X线照射,DR4和DR5mRNA表达水平均显著高于正常对照组(P<0.05),但只有DR5mRNA表达水平显著高于6Gy X线照射组(P<0.05)。Westernblotting结果显示,与正常对照组比较,DR4和DR5蛋白在pEgr1-hsTRAIL组无明显变化,而在6Gy X线照射和pEgr1-hsTRAIL+6Gy X线照射组表达增加,其中DR5蛋白在pEgr1-hsTRAIL+6Gy X线照射组增加更明显。结论:重组表达质粒pEgr1-shTRAIL能够增强A549细胞放射敏感性,且能增强DR5辐射诱导表达,而对DR4的辐射诱导表达无明显增强作用。Objective To measure the changes of the radiosensitivity in human lung adenocarcinoma A549 cells transfected with pEgr1-hsTRAIL plasmid and the effect on death receptor（DR） 4 and DR5 expressions,and to explore the radiosensitizing effect of pEgr1-hsTRAIL plasmid and possible mechanism on inducing apoptosis.Methods There were normal control,pEgr1-hsTRAIL,6 Gy X-rays,and pEgr1-hsTRAIL ＋ 6 Gy X-rays groups in the experiment.After the A549 cells were transfected with liposome,and irradiated with X-rays,colony formation assay was used to measure the radiosensitivity,and reverse transcription PCR（RT-PCR） was performed to detect the DR4 and DR5 mRNA expressions,and Western blotting was applied to determine the DR4 and DR5 protein expressions.Results The D0 values of A549 cells in normal control group and pEgr1-hsTRAIL group were 3.26 and 1.91 Gy,respectively,it indicated that pEgr1-hsTRAIL plasmid could enhance the radiosensitivity in A549 cells.The RT-PCR results showed that as compared with normal control group,the DR4 and DR5 mRNA expression levels in pEgr1-hsTRAIL group had no significant change,but those in 6 Gy X-rays group were increased significantly（P0.05）,and those in pEgr1-hsTRAIL ＋ 6 Gy X-rays group were also increased significantly（P0.05）;the DR5 mRNA expression level in pEgr1-hsTRAIL ＋ 6 Gy X-rays group was higher than that in 6 Gy X-rays group（P0.05）.The Western blotting results showed that the DR4 and DR5 protein expressions in pEgr1-hsTRAIL group did not change obviously compared with normal control group,but those in 6 Gy X-rays and pEgr1-hsTRAIL ＋ 6 Gy X-rays groups were increased,and the DR5 protein expression in pEgr1-hsTRAIL ＋6 Gy X-rays group was increased mostly.Conclusion The recombinant plasmid pEgr1-hsTRAIL can enhance the radiosensitivity of A549 cells,and has the enhancing effect on DR5 expression induced by radiation,but no same effect on DR4 expression.

关 键 词：早期生长反应因子1 肺腺癌 人分泌型肿瘤坏死因子相关凋亡诱导配体 辐射敏感性 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]