C-erbB-2 shRNA对小鼠肺腺癌细胞化疗敏感性的影响及其机制  

Effect of C-erbB-2 shRNA on chemosensitivity of mouse lung adenocarcinoma cells and its mechanism

作　　者：曹新梅[1] 张岱权[2] 王栩[1] 夏纪毅[3] 黄黎[1] 高燕[1] 

机构地区：[1]泸州医学院免疫学教研室,四川泸州646000 [2]泸州医学院附属医院中医科,四川泸州646000 [3]泸州医学院药物与功能性食品研究中心,四川泸州646000

出　　处：《吉林大学学报（医学版）》2014年第4期777-781,共5页Journal of Jilin University：Medicine Edition

基　　金：四川省卫生厅科研基金资助课题(100236)

摘　　要：目的:探讨人表皮生长因子受体2短发夹RNA(C-erbB-2shRNA)对小鼠肺腺癌Lewis细胞化疗敏感性的影响及其机制,为非小细胞肺癌尤其是腺癌的治疗提供新的思路。方法:常规培养小鼠肺腺癌Lewis细胞,分为未转染组、pGPU6/RFP/Neo-shNC组和pGPU6/RFP/Neo-erbB-2组;应用Lipofectamine 2000将质粒快速转染各组Lewis细胞;应用RT-PCR法检测各组细胞C-erbB-2mRNA表达水平;应用Western blottting法检测各组细胞C-erbB-2蛋白表达水平。将Lewis细胞分为未转染对照组、pGPU6/RFP/Neo-shNC组、卡铂组、pGPU6/RFP/Neo-erbB-2组、pGPU6/RFP/Neo-erbB-2+卡铂组和pGPU6/RFP/Neo-shNC+卡铂组;应用流式细胞术检测各组细胞凋亡率;应用Western blotting法检测各组细胞Bcl-2和Bax蛋白表达水平。结果:pGPU6/RFP/Neo-erbB-2组C-erbB-2mRNA和蛋白表达水平均低于未转染组和pGPU6/RFP/Neo-shNC组;pGPU6/RFP/Neo-erbB-2+卡铂组细胞凋亡率高于其他各组(P<0.01);与其他各组比较,pGPU6/RFP/Neo-erbB-2+卡铂组Bcl-2蛋白表达水平下降、Bax蛋白表达水平增加。结论:C-erbB-2shRNA能增强小鼠肺腺癌Lewis细胞对卡铂的敏感性,其机制可能是通过上调Bax蛋白、下调Bcl-2蛋白表达,进而增强卡铂诱导的细胞凋亡。Objective To investigate the effect of C-erbB-2 shRNA on chemosensitivity of mouse lung adenocarcinoma cells and its mechanism,and to find new therapy method for non-small cell lung cancer,especial lung adenocarcinoma.Methods The mouse lung adenocarcinoma Lewis cells were cultured regularly and divided into non-transfected group,pGPU6/RFP/Neo-shNC group and pGPU6/RFP/Neo-erbB-2group.The plasmids were synthesized and transfected into Lewis cells in each group by Lipofectamine 2000.The expression levels of C-erbB-2mRNA and protein in the cells in various groups were tested by RT-PCR and Western blotting method,respectively.The Lewis cells were divided into non-transfected group,pGPU6/RFP/Neo-shNC group,carboplatin group,pGPU6/RFP/Neo-erbB-2group,pGPU6/RFP/Neo-shNC＋carboplatin group and pGPU6/RFP/Neo-erbB-2＋carboplatin group.The apoptotic rates of the cells in each group were detected by flow cytometry;the expression levels of Bcl-2and Bax proteins in each group were determined by Western blotting method.Results The expression levels of C-erbB-2mRNA and protein in pGPU6/RFP/Neo-erbB-2group were lower than those in non-transfected group and pGPU6/RFP/Neo-shNC.The apoptotic rate of the cells in pGPU6/RFP/Neo-erbB-2＋carboplatin group was the highest in all of the groups（P〈0.01）;compared with the others,the expression of Bax protein in pGPU6/RFP/Neo-erbB-2＋carboplatin group was increased,while the expression level of Bcl-2protein was decreased.Conclusion C-erbB-2shRNA can increase the Lewis cells＇ sensitivity to carboplatin.The mechanism may be that it can enhance the Lewis cells＇ apoptosis induced by carboplatin through increasing the expression of Bax protein and decreasing the expression of Bcl-2protein.

关 键 词：人表皮生长因子受体2 短发夹RNA 肺腺癌细胞 化学疗法 

分 类 号：R734.2[医药卫生—肿瘤]