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目的探讨细胞膜与细胞骨架连接蛋白埃兹(Ezrin)对人脐静脉内皮细胞生长和转移能力的影响。方法培养人脐静脉内皮细胞,选择3对小干扰RNA(siRNA)序列下调Ezrin的表达,实时定量PCR和Western-blot印迹法分别检测Ezrin mRNA和蛋白表达水平的变化。BrdU ELISA法检测Ezrin siRNA干扰后细胞增殖能力的改变,伤口愈合实验检测Ezrin siRNA干扰后细胞迁移能力的变化,流式细胞术检测Ezrin siRNA干扰后细胞凋亡能力。结果实时定量PCR和Western-blot印迹结果显示,siRNA干扰后Ezrin的基因和蛋白水平均显著下调;BrdU法显示Ezrin siRNA干扰后细胞生长速率降低(P<0.05),划痕实验显示Ezrin siRNA干扰后细胞迁移率降低(P<0.05),流式细胞仪检测显示Ezrin siRNA干扰后细胞凋亡率升高。结论 Ezrin蛋白可影响脐静脉内皮细胞的增殖和迁移能力,推测其在血管功能和肿瘤研究中可能具有重要意义。
Objective To investigate the effects of cell membrane and cytoskeletal connexin Ezrin on the growth and metastasis of human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells were cultured and three siRNAs were selected to downregulate the expression of Ezrin. The mRNA and protein levels of Ezrin were detected by real-time PCR and Western-blot respectively. Ezrin siRNA was used to detect the proliferation of Ezrin siRNA. BrdU ELISA was used to detect the changes of cell migration after Ezrin siRNA treatment. The apoptosis of Ezrin siRNA was detected by flow cytometry. Results The real-time quantitative PCR and Western blotting results showed that Ezrin gene and protein levels were significantly down-regulated after siRNA interference. The BrdU method showed that the cell growth rate was decreased after Ezrin siRNA interference (P <0.05) Cell migration was decreased (P <0.05). Flow cytometry showed that the apoptosis rate of Ezrin siRNA was increased after interference with siRNA. Conclusion Ezrin protein can affect the proliferation and migration of human umbilical vein endothelial cells, suggesting that it may play an important role in vascular function and tumor research.