siRNA干扰Pokemon基因对鼻咽癌CNE-2细胞生长作用的研究  

Influence of silencing Pokemon genes using small-interfering RNA on growth of CNE-2 cells of NPC

作　　者：牛道立[1,2] 李建锋[2] 何芬[2] 邹俊涛[3] 周琼芳[2] 魏璇[2] 李杨[2] 陈龙华[1] 

机构地区：[1]南方医科大学附属南方医院放疗科,广东广州510515 [2]广州医学院第一附属医院放疗科,广东广州510120 [3]中山大学中山医学院人体解剖学教研室,广东广州510080

出　　处：《中华肿瘤防治杂志》2012年第14期1041-1045,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81000545);广东省自然科学基金(9451008901002145;S2011010003915);广东省医学科研基金(B201-0066);广州市教委重大科研项目(61100);广州市教育局科技计划项目(08A040);广州市医药卫生科技项目(201102A213119);广州市科技计划项目(10C32060117);教育部高等学校博士学科点专项科研基金(20090171120047)

摘　　要：目的:探讨siRNA干扰Pokemon基因对鼻咽癌CNE-2细胞生长作用的影响。方法:利用siRNA预测软件得到3条siRNA编码DNA序列,并制备合成siRNA片段,利用RT-PCR和蛋白质印迹法检测其沉默Pokemon基因的效果,筛选出沉默效果最佳的siRNA片段。利用MTT法和流式细胞术检测CNE-2细胞生长与凋亡情况。结果:筛选出一条≥50%沉默效应的siRNA片段,转染鼻咽癌CNE-2细胞后,Pokemon基因表达明显下降(P<0.05),p14ARF和p53基因表达上调,P<0.05。鼻咽癌CNE-2细胞增殖受到抑制(P=0.015),凋亡率升高,P=0.008。结论:Pokemon基因沉默后可抑制CNE-2细胞生长,促进细胞凋亡,其机制可能与p14ARF和p53的表达上调有关。OBJECTIVE： To construct recombinant plasmids containing microRNA targeting the Pokemon gene and investigate the effects of miRNA-mediated downregulation of the Pokemon gene on the proliferation of CNE-2 cells. METHODS： Three siRNA were designed using siRNA prediction software. The best one was chosen according to the result of Pokeman gene silencing through RT-PCR and Western blot. The expression of ARF and p53 gene was observed, and then the influence of CNE-2 cell growth and apotosis were detected by MTT and flow cytometry. RESULTS： After silencing Pokeman gene, the expression of Pokeman gene of nasopharyngeal carcinoma CNE-2 cell was obviously downregulated （P〈0.05） ,but the expression of p14ARF and p53 gene was significantly increased （P〈0.05）. The growth of CNE-2 cell was suppressed （P=0. 015） and its apoptosis was triggered （P=0. 008）. CONCLUSIONS：The results show that the growth of CNE-2 cell is inhibited and the apoptosis is promoted when Pokemon gene is silenced. The mechanism of this phenomenon may related to the upexpression of p14ARE and p53 gene.

关 键 词：RNA干扰 基因表达 鼻咽肿瘤 转染 

分 类 号：R739.63[医药卫生—肿瘤]