RNAi沉默Survivin基因抑制人鼻咽癌细胞生长  

The cell growth inhibition induced by RNA interference to silence Survivin gene in human nasopharyngeal carcinoma

作　　者：武炜[1,2] 申咏梅[1,2] 吾为一[1] 黄宏[1] 石怡珍[1,2] 王巍[1] 荣光[1] 于喜龙[1,2] 倪凯茹[1] 

机构地区：[1]苏州大学附属第二医院核医学科,江苏苏州215004 [2]苏州大学附属第二医院临床检验教研室,江苏苏州215004

出　　处：《苏州大学学报（医学版）》2012年第3期331-335,共5页Suzhou University Journal of Medical Science

基　　金：苏州市科技发展计划(应用基础研究)项目(SYSD2010118)

摘　　要：目的观察RNA干扰沉默Survivin基因对人鼻咽癌CNE2细胞在体内外生长的抑制作用。方法将Survivin特异性shRNA表达载体pGenesil-1-survivin经LipofectamineTM2000介导转染人鼻咽癌CNE2细胞,经G418筛选获得稳定转染的CNE2细胞,荧光显微镜和流式细胞仪检测细胞转染效率;逆转录-聚合酶链反应(RT-PCR)和Western blot分别检测Survivin基因mRNA和蛋白表达水平;噻唑蓝(MTT)法检测细胞增殖能力的变化。以稳定转染细胞建立裸鼠荷瘤模型,观测瘤体积的变化,绘制移植瘤生长曲线。结果流式细胞仪检测pGenesil-1-survivin细胞转染百分率为(87.13±2.21)%;RT-PCR和Western blot结果显示,pGenesil-1-survivin组对人鼻咽癌CNE2细胞Survivin mRNA和蛋白的抑制率分别为64.55%和51.24%,均较随机序列构建载体的阴性对照(pGenesil-1-NC)组和空白对照组明显下调(均P<0.05);MTT结果显示,pGenesil-1-survivin组细胞增殖能力明显较pGenesil-1-NC组和空白对照组降低(均P<0.05);体内实验表明,pGenesil-1-survivin组较pGenesil-1-NC组和空白对照组能有效抑制BALB/C裸鼠人鼻咽癌的生长(均P<0.05)。结论沉默Survivin基因可以有效抑制人鼻咽癌CNE2细胞在体内外的增殖,Survivin基因有望成为鼻咽癌基因治疗的有效靶点。Objective To investigate the inhibition effect of RNA interference targeting survivin gene of human nasopharyngeal carcinoma CNE2 ceils in vitro and in vivo. Methods pGenesil-l-survivin ex- pression vector of short hairpin RNA （shRNA） specific to survivin gene was transfeeted into human naso- pharyngeal carcinoma CNE2 cells through LipofectamineTM2000 reagent. Stably transfected CNE2 cells were obtained after G418 screening and detected by fluorescence microscope and flow cytomytry （FCM） to analyze the cell transfection efficiency. The mRNA and protein expression of survivin were determined by reverse transcription-polymerase chain reaction （RT-PCR） and FCM, respectively. Cell proliferation ability was determined by methyl thiazol tetrazolium （MTT） assay. BALB/c nude mice bearing stably transfeeted human nasopharyngeal carcinoma CNE2 cells were established. Tumor sizes were measured and tumor growth curves were made. Results FCM results showed that positive transfected percentage of pGenesil-l-survivin was （87.13±2.21 ） %. The mRNA and protein expression of survivin inhibited by 64.55% and 51.24% respectively and deduced significantly when compared with that of either pGenesil-1-NC （ expression vector with scrambled small interfering RNA） transfected ceils or control ceils （P 〈 0.05）. The MTI＂ results showed that the proliferation rate of CNE2 ceils stably transfected with pGenesil- 1-survivin was significantly reduced when compared with that of either pGenesil-l-NC group or control group （P 〈 O. 05 ）. In vivo mice bearing tumor, pGenesil-1 osurvivin showed a significant inhibitory effect on the growth of transplanted human nasopharyngeal carcinoma, resulting in a statistically significant difference compared with the other groups （P 〈 0.05 ）. Conclusion Survivin gene silence can effective- ly inhibit the proliferation ability of human nasopharyngeal carcinoma CNE2 ce｜ls in vitro and in vivo. Sur- vivin gene may be a promising target in the gene therapy of human naso

关 键 词：RNAI SURVIVIN pGenesil-1-survivin 鼻咽癌 CNE2细胞 

分 类 号：R739.63[医药卫生—肿瘤]