小干扰RNA靶向抑制埃兹蛋白对肠癌细胞生物学行为的影响  

The influence of silencing Ezrin expression by RNA interference on the biological behaviors of colorectal carcinoma in vitro

作　　者：颜歌[1] 周晓燕[1] 邹丽萍[1] 杨银龙[1] 祝蕾[1] 罗建民[1] 杜祥[1] 

机构地区：[1]复旦大学肿瘤医院病理科,复旦大学上海医学院肿瘤学系,复旦大学大肠癌诊治中心,上海200032

出　　处：《肿瘤》2008年第1期21-24,共4页Tumor

基　　金：上海市科学技术发展基金资助项目(编号:024119010,064119512)

摘　　要：目的:探讨细胞膜与细胞骨架连接的埃兹蛋白(Ezrin)对肠癌细胞生长和侵袭转移能力的影响。方法:选取肠癌细胞系(Lovo,SW480)作为研究材料,采用小干扰RNA(small interfering RNA,siRNA)方法下调Ezrin的表达,分别通过实时荧光定量PCR和Western印迹法检测siRNA对Ezrin mRNA和蛋白表达水平的下调及下调作用随干扰时间的变化。根据Western印迹法检测结果,选取Ezrin蛋白表达最低的时间点检测干扰后肠癌细胞生物学行为的改变。运用CCK-8法检测2株细胞干扰前后增殖能力的改变,流式细胞仪检测细胞周期和细胞凋亡,Transwell实验检测肠癌细胞侵袭和转移能力的变化。结果:实时荧光定量PCR和Western印迹结果显示,Ezrin siRNA对Ezrin mRNA和蛋白表达水平均有显著的下调作用。下调Ezrin蛋白后,2株肠癌细胞S期细胞比例略有下降,G1期细胞比例略有上升,与对照组比差异无统计学意义(P>0.05);2株细胞凋亡指数增加,与对照组比差异有统计学意义(P<0.05);肿瘤细胞的运动侵袭能力下降,穿过人工基底膜的细胞数量明显减少,与干扰前比差异有统计学意义(P<0.01)。结论:Ezrin在肠癌生长和侵袭转移过程中发挥重要作用,有可能成为抑制肠癌发生发展的新靶点。Objective： To study the role of membrane-cytoskeleton linker Ezrin in the growth and metastasis of human colorectal carcinoma （CRC） cell lines. Methods： Human CRC cell lines Lovo and SW 480 were cultured. Three pairs of small interfering RNA （siRNA） targeting Ezrin were transfected into the CRC cells. Western blotting was used to detect the transfection rates so as to screen the most effective siRNA which had been transfected into the CRC cells. Real-time PCR and Western blotting were used to detect the downregulation of Ezrin mRNA and protein at different time points. We observed the biological behaviors of CRC cells when the expression level of Ezrin was the lowest. CCK-8 method was used to detect the proliferation of the cells. Flow cytometry was used to detect the cell cycle and apoptosis. Transwell test was used to detect the invasion and migration ability of the transfected CRC cells. Results ： Real- time PCR and Western blotting results revealed that Ezrin siRNA notably down-regulated Ezrin expression at both mRNA and protein levels. Down-regulation of Ezrin expression distinctly decreased the proliferation rates of the two cell lines. After RNAi treatment, the cell proportion in S phase decreased slightly and the cell proportion in G1 phase increased slightly in the two cell lines （ P 〉 0.05 ）. The apoptotic index increased in the three siRNA transfected groups compared with the control group （ P 〈 0.05 ）. The migration and invasion ability of CRC cells significantly decreased after siRNA interference （ P 〈 0.01 ）. Conclusion ： Ezrin plays an important role in the process of proliferation, migration, and invasion of CRC cells. It has the potential to become a new target to inhibit tumorigenesis and development of CRC.

关 键 词：肠肿瘤 RNA干扰 基因表达 

分 类 号：R735.3[医药卫生—肿瘤]