siRNA下调EZH2基因对肾癌细胞系769-P增殖的影响  

Effect of EZH2 after Silenced by siRNA on the Proliferation of Renal Cell Carcinoma Cell Line 769-P

作　　者：王国栋[1,2] 党亚正[1] 魏铭[2] 郑甲[2] 温伟红[2] 

机构地区：[1]解放军第323医院肿瘤中心 [2]第四军医大学肿瘤生物学国家重点实验室,陕西西安710032

出　　处：《现代生物医学进展》2014年第23期4440-4443,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81171924)

摘　　要：目的:运用小干扰RNA下调果蝇zeste基因增强子人类同源物(enhancer ofzeste homolog 2,EZH2)在肾癌细胞系769-P中的表达,明确其对肾癌细胞增殖的影响。方法:将处于对数生长期769-P细胞分为实验组(experiment group)、阴性对照组(negative group)、空白对照组(blank group),合成靶向EZH2基因的小干扰RNA片段(EZH2-siRNA)和无效序列片段后,通过脂质体介导分别转染至实理组和阴性对照组,空白对照组未做任何处理。以qRealtime-PCR检测EZH2基因mRNA水平的变化情况,以MTT法检测各组细胞增殖变化;流式细胞术(FCM)检测转染后细胞周期变化情况。结果:实理组中EZH2在mRNA表达水平明显受抑制;MTT实验中第4天始,实验组中769-P细胞的增殖能力开始受抑制,第5天时实验组细胞抑制更明显,与阴性对照组和空白组比较差异有统计学意义(P<0.05)。siRNA转染后实验组中G0/G1期细胞比例明显增多(81.32±3.14)%,与阴性对照组(44.13±1.52)%和空白对照组(45.71±2.32)%差异有统计学意义。结论:EZH2-siRNA可有效下调并抑制肾癌细胞769-P的增殖,EZH2在肾癌的发生、发展中发挥了重要作用,为下一步研究肾癌基因治疗提供了理论支持。Objective： To investigate the influence of siRNA targeting the enhancer of homolog2（EZH2） gene on the proliferation of renal cell carcinoma（RCC） cell line 769-P in vitro. Methods： The cell line 769-P which were in the period of logarithmic phase were divided into experiment group, negative group and blank group. siRNA sequence targeting EZH2 were designed and synthesized according to genebank, with a scramble sequence as negative control, which were transefected to the 769-P cells conducted by Lipofectamine 2000. The expression of EZH2 in mRNA level was detected by qRT-PCR after downregulated. MTT assay was used to observe the growth of 769-P cells in experiment group, negative group and blank group. And, cell cycle distribution of 769-P cells were detected by flow cytometry. Results： The expression of EZH2 in mRNA level was significantly suppressed by siRNA. MTT assay results revealed that cell inhibitory rate of experiment group was more lower than that of negative group and blank group. The proliferation of769-P cells were inhibited remarkably in experiment group with cell rate of G0/G1 phrase being（81.32±3.14）%, and there were statistic difference as compared with negative control group（44.13±1.52）% and blank group（45.71±2.32）%. Conclusions： EZH2 gene silencing can be conducted effectively by siRNA. EZH2 may play important roles in the development and progression of RCC, EZH2 may be a potential theraputic target, and provides an important theoretical basis for gene therapy of RCC.

关 键 词：肾细胞癌 RNAI EZH2 

分 类 号：R737.11[医药卫生—肿瘤]