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目的研究TALEN(transcription activator-like effector nuclease)靶向敲除REST(repressor element 1-silencing transcription factor)对U-87细胞增殖和迁移能力的影响。方法使用分子克隆手段构建TALEN质粒;通过western blot分析TALEN打靶效率;分别使用CCk-8法和Transwell小室法检测细胞增殖和迁移能力。结果 TALEN能够有效打靶U-87细胞中的REST,蛋白水平显著下调;TALEN靶向敲除REST能够显著降低U-87细胞的增殖和迁移水平。结论 TALEN能够高效打靶U-87细胞中的REST,进而抑制U-87细胞的增殖和迁移能力。
Objective To investigate the effect of repressor element 1-silencing transcription factor (TALEN) on the proliferation and migration of U-87 cells. Methods TALEN plasmid was constructed by molecular cloning. TALEN targeting efficiency was analyzed by western blot. Cell proliferation and migration were detected by CCk-8 assay and Transwell chamber assay. Results TALEN could effectively target REST in U-87 cells, and the protein level was significantly down-regulated. TALEN-targeted knockdown of REST significantly reduced the proliferation and migration of U-87 cells. Conclusion TALEN can efficiently target REST in U-87 cells and inhibit the proliferation and migration of U-87 cells.