shRNA—CXCR4对人胃癌BGC823细胞增殖迁移的影响  

Inhibitory effects of shRNA-CXCR4 on proliferation and migration of human gastric cancer Bgc823 cells

作　　者：朱志强[1] 宁忠良[1] 胡斌[1] 汤志刚[1] 

机构地区：[1]安徽医科大学附属省立医院普外科,合肥230001

出　　处：《中华实验外科杂志》2011年第11期1899-1902,共4页Chinese Journal of Experimental Surgery

摘　　要：目的构建针对人CXCR4基因的shRNA干扰表达载体，观察CXCR4基因表达对人胃癌细胞株BGC823细胞增殖和迁移能力的影响。方法根据人CXCR4序列设计并构建CXCR4基因的shRNA真核表达载体，以脂质体转染的方法将CXCR4干扰质粒转染至人胃癌细胞株BGC823，经G418筛选出稳定转染的阳性细胞，应用流式细胞仪技术检测其细胞周期变化，噻唑蓝（MTT）比色法检测CXCR4干扰对胃癌细胞增殖的影响，采用Trans—well小室法检测细胞体外迁移能力。结果经测序证明CXCR4的shRNA基因已成功插入Psilencer4．1质粒中，构建的干扰质粒能够显著抑制CXCR4的表达，CXCR4mRNA被抑制了（2．7±0．2）倍（P〈0．01）。CXCR4基因的表达下调将胃癌细胞阻滞在G1期（由84．2％升至94．1％），MTT法检测CXCR4-shRNA干扰细胞增殖（P〈0．05），且穿过Trans—well小室膜的细胞数显著下降（P〈0．05）。结论干扰CXCR4的表达可显著抑制胃癌细胞的增殖和迁移能力。Objective To construct the shRNA against human CXCR4 gene expression vector interference observed inhibition of gastrin gene expression in human gastric cancer cell line BGC823 cell proliferation and migration capacity. Methods The human CXCR4 sequence designed and constructed human gastrin gene eukaryotic expression vector, liposome transfection method gastrin interference plasmid into human gastric cancer cell lines BGC823, by G418 screened positive for a stable transfected cells. Flow cytometry to detect the cell cycle changes. Methyl thiazol tetrazolium （MTF） assay interference CXCR4 in gastric cancer cell proliferation. The use of Trans-well chamber assay in vitro migration. Results The sequence proved the CXCR4 gene was successfully inserted Psilencer4. 1 shRNA plasmid to construct the interference plasmid can significantly inhibit the expression of CXCR4, the expression of CXCR4mRNA was inhabited 2. 7 ±0. 2 times than it in the control group （P 〈 0. 01 ）. CXCR4 gene expression down to gastric cancer cells were arrested in G1 phase（from 84. 2% up to 94. 1% ）, MTT CXCR4-shRNA detected by interfering with cell proliferation （P 〈 0. 05 ）, and the Trans-well chamber through the membrane of cells was significantly decreased （P 〈 0. 05）. Conclusion The expression of CXCR4 interference significantly inhibited gastric cancer cell proliferation and migration.

关 键 词：胃癌 增殖 SHRNA CXCR4 

分 类 号：R735.2[医药卫生—肿瘤]