rna干扰foxm1基因抑制食管鳞癌细胞增殖 精灵论文.docVIP

RNA 干扰 FOXM1 基因抑制食管鳞癌细胞增殖 管国波，刘玮婧，戚玺君，王大巾，章小斌，赵辅昆，丁明 （浙江理工大学生命科学学院分子酶学与蛋白质组学研究室，杭州 310018） 摘要：目的：利用 RNA 干扰技术下调食管鳞癌细胞 TE-1 中 Fockhead box M1（FOXM1） 的表达水平，探讨 FOXM1 对食管鳞癌细胞增殖和迁移能力的影响。方法：构建 3 组针对 FOXM1 基因不同位点的 shRNA 表达载体（pLKO.1-FOXM1 shRNA 1，pLKO.1-FOXM1 shRNA 2，pLKO.1-FOXM1 shRNA 3），与病毒包装质粒（pCMV-DR8.2，pCMV-VSV-G） 共转染 HEK293T 细胞，包装病毒后侵染食管鳞癌 TE-1 细胞，利用嘌呤霉素筛选稳定株。 Western blot 检测稳定株 FOXM1 的表达水平，利用 MTT 实验和划痕实验分析 FOXM1 表达 水平下调对 TE-1 细胞增殖能力和迁移能力的影响。结果：测序结果表明成功构建了 3 个 FOXM1 干扰载体；Western blot 检测显示 3 个干扰载体抑制 FOXM1 表达的能力不同，其中 RNA 干扰组 pLKO.1-FOXM1 shRNA 2 的抑制效果最好；MTT 和划痕实验结果表明 RNA 干 扰组细胞的增殖能力和迁移能力有所下降。结论：体外实验证实下调 FOXM1 的表达可抑制 食管鳞癌细胞的增殖和迁移，这为进一步体内研究 FOXM1 在食管鳞癌发生和转移中的作用 提供了理论和实验依据。 关键词： 食管鳞癌；Fockhead box M1；RNA 干扰；细胞增殖；细胞迁移 中图分类号：Q513 Down-regulation of FOXM1 by RNA Interference Inhibits Proliferation of Esophageal Squamous Cell Carcinoma Cells Guan Guobo, Liu Weijing, Qi Xijun, Wang Dajin, Zhang Xiaobin, Zhao Fukun, Ding Ming (School of Life Sciences, Zhejiang Sci-Tech University, HangZhou 310018) Abstract: Objective: To investigate the effects of Fockhead box M1 (FOXM1) on the proliferation and migration of ESCC cells, RNAi was performed to specifically knock down FOXM1 expression in ESCC cells, TE-1. Methods: Three shRNA plasmids specifically targeting different sites of FOXM1 were constructed and transfected into HEK293T cells with pCMV-DR8.2, pCMV-VSV-G, generating lentiviral particles and infected the target cell. Cells that stably express shRNA were selected by puromycin. FOXM1 expression was measured by Western blot. The effects of FOXM1 shRNA on cell proliferation and cell migration were examined by methyl thiazolyl tetrazolium (MTT) and wound-healing assay. Results: Three recombinant plasmids targeting FOXM1 were constructed successfully. The inhibitory effects of 3 shRNA plasmids varied, pLKO.1-FOXM1 shRNA 2 group had the most best effect. As compared to pLKO.1-scramble group, the pLKO.1-FOXM1 shRNA 2 group showed lower proliferation. Further