RNA干扰抑制c-jun基因表达对放射抗拒人鼻咽癌细胞系CNE-2R放射敏感性的影响  

Effect of c-jun gene RNAi on the radiosensitivity of radioresistant human nasopharyngeal carcinoma cell line CNE-2R

作　　者：郭思言[1] 朱小东[1] 葛莲英[1] 曲颂[1] 李龄[1] 郭亚[2] 梁世雄[1] 苏芳[1] 

机构地区：[1]广西医科大学附属肿瘤医院广西壮族自治区肿瘤防治研究所放疗科区域性高发肿瘤早期防治研究教育部重点实验室(广西医科大学),南宁530021 [2]西安交通大学第二附属医院肿瘤科

出　　处：《中华放射医学与防护杂志》2014年第12期895-899,共5页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金（30860329）;广西壮族自治区自然科学基金（桂科自0832229）;广西高等学校重大科研资助项目（201101ZD004）

摘　　要：目的 利用RNA干扰技术抑制放射抗拒人鼻咽癌细胞系CNE-2R中高表达的c-jun基因,研究其对CNE-2R细胞放射敏感性的影响及其潜在机制.方法 设计合成3组特异性针对c-jun基因的siRNA,构建慢病毒vshRNA载体,感染CNE-2R细胞,采用RT-PCR和Western blot方法检测各组对目的基因的抑制效果;并通过克隆形成实验测定其放射敏感性,CCK-8实验检测细胞的存活能力,流式细胞术检测细胞凋亡率.结果 重组慢病毒c-jun-RNAi-LV2能明显下调CNE-2R细胞中c-jun的mRNA和蛋白水平的表达（F =217.20、42.07,P＜0.05）.6MVX射线联合c-jun-RNAi-LV2组在0、2、4、6和8 Gy照射后细胞的吸光度值均显著低于未转染组和阴性对照组（F=42.70 ～ 200.67,P＜0.05）.克隆形成实验显示,c-jun-RNAi-LV2组与未转染组及阴性对照组相比,SF2、D0和Dq值均减小,放射增敏比（SERD0）为1.41.细胞凋亡实验结果显示,未经照射的c-jun-RNAi-LV2组与联合2 Gy照射的凋亡率分别为（20.93±1.99）％和（38.17±0.83）％,均高于未转染组和阴性对照组的凋亡率[0 Gy：（10.97±0.70）％和（20.43±0.25）％;2 Gy：（10.80±1.25）％和（19.53±1.50）％;F=50.54、330.14,P＜0.05].结论 RNA干扰技术可有效抑制放射抗拒人鼻咽癌细胞系CNE-2R中高表达的c-jun基因,使细胞放射敏感性增高,其作用可能与c-jun基因的下调使细胞增殖能力减弱,细胞凋亡增多有关.Objective To investigate the effect of c-jun RNAi on the radiosensitivity of radioresistant human nasopharyngeal carcinoma cell line CNE-2R and the possible mechanisms.Methods Three siRNA sequences specific for c-jun gene were designed,and lentvirus vector with c-jun shRNA was constructed and infected into CNE-2R.The gene silencing efficiency of these recombinants was detected by RT-PCR and Western blot.Radiosensitivity,cell proliferation and apoptosis were measured using colony formation assay,CCK-8 assay and flow cytometry respectively.Results Recombinant lentiviral c-jun-RNAi-LV2 obviously inhibited c-jun expression at mRNA and protein level （F =217.20,42.07,P 〈 0.05）.The cell proliferation of combination group （c-jun-RNAi-LV2 plus 0,2,4,6,8 Gy of 6 MV X-rays） was significantly lower than that of the other two groups （F =42.70-200.67,P 〈0.05）.The cloning formation of c-jun-RNAi-LV2 group was significantly lower than that of non-transfected group and NC-transfected group and its sensitizing enhance rate （SER） at D0was 1.41.The apoptosis rates of 0,2 Gy X-ray radiation in c-jun-RNAi-LV2 group,non-transfected group and NC-transfected group were （20.93 ±1.99）%,（38.17 ±0.83）% ;（10.97 ±0.70）%,（20.43 ±0.25）% ;（10.80 ±1.25）% and （19.53±1.50）% （F =50.54,330.14,P 〈0.05）,respectively.Conclusions RNA interference inhibited the c-jun expression and enhanced the radiation sensitivity of human nasopharyngeal carcinoma cell line CNE-2R,perhaps due to that the c-jun-RNAi-LV2 mediated c-jun down-regulation inhibited cell proliferation and promoted cell apoptosis.

关 键 词：鼻咽癌 放射抗拒 C-JUN基因 RNA干扰 

分 类 号：R739.63[医药卫生—肿瘤]