【摘要】 胃癌细胞分泌的胃泌素与胃癌的发生、发展密切相关.为了探讨胃泌素对胃癌细胞增殖、迁移和侵袭的影响,本文构建靶向胃泌素基因的siRNA表达载体,转染胃癌细胞AGS,成功获得沉默胃泌素基因的稳转胃癌细胞株AGS/Gas-siRNA.用MTT实验、软琼脂集落形成实验、细胞伤愈实验、Transwell实验及ELISA检测沉默胃泌素基因后细胞的增殖、迁移、侵袭及转移相关蛋白基质金属蛋白酶-2(MMP-2)和血管内皮生长因子(VEGF)的含量.结果显示:与空载体转染的对照细胞比较,沉默胃泌素基因的细胞,其增殖率和克隆形成率显著降低,迁移和侵袭到Transwell下室的细胞数分别降低了31.6%和34%.培养上清液中MMP-2和VEGF含量也低于对照细胞.结果提示,沉默胃泌素基因的胃癌细胞,通过降低MMP-2和VEGF分泌,抑制了细胞的增殖、迁移和侵袭,这可能是胃泌素促进胃癌侵袭转移的机制之一.更多还原

【Abstract】 Secreted by gastric cancer cells,gastrin was closely related to gastric cancer development and progression. To study the effects of gastrin on cell proliferation,migration and invasion,we constructed a eukaryotic siRNA expression vector to knock down gastrin and transfected into AGS cells. A stable gastrin-silenced cell line AGS /Gas-siRNA was obtained. MMT,colony formation,wound healing and Transwell assay were performed. The concentrations of matrix metalloproteinases-2( MMP-2) and vascular endothelial growth factor( VEGF) in culture medium were measured by ELISA. The results showed that the measurements of proliferation and colony formation in soft agar were significantly decreased in AGS /Gas-siRNA cells as compared to the control cells. The cells in the lower chamber of Transwell were reduced by 31. 6 % to 34 %. The medium levels of MMP-2 and VEGF were also reduced. These suggested that the silencing of gastrin reduced the secretion of MMP-2 and VEGF to inhibit cell proliferation,migration and invasion.更多还原