稳定转染15-PGDH基因对人胃癌细胞生长和迁移的影响  

Effect of Stable Transfection of 15-PGDH Gene on Growth and Migration of Human Gastric Cancer Ceils

作　　者：申玉翠 靖大道[1] 叶赛[2] 王弢[2] 张庆华[2] 

机构地区：[1]上海交通大学附属第一人民医院消化科,200080 [2]生物芯片上海国家工程研究中心

出　　处：《胃肠病学》2009年第5期274-278,共5页Chinese Journal of Gastroenterology

摘　　要：背景：前期研究表明胃癌组织中NAD＋依赖性15-羟基前列腺素脱氢酶（15-PGDH）表达明显减低，瞬时转染15-PGDH对人胃癌细胞的生长和迁移有一定抑制作用。目的：建立稳定转染15-PGDH基因的人胃癌细胞株SGC7901．观察恢复15-PGDH表达对胃癌细胞生长和迁移的影响，探讨15-PGDH与胃癌发生、发展的关系。方法：以双酶切和质粒测序鉴定真核表达质粒pcDNA3／15-PGDH，采用脂质体法将质粒转染人SGC7901细胞，G418筛选稳定转染株．实时聚合酶链反应（PCR）和蛋白质印迹法鉴定。分别以甲基噻唑基四唑（MTT）实验和细胞划痕实验检测稳定转染15-PGDH基因的SGC7901细胞株的增殖情况和迁移能力。结果：经4周G418筛选以及实时PCR和蛋白质印迹法鉴定，得到4株可稳定、较高水平表达15-PGDH的SGC7901细胞株。稳定转染15-PGDH基因的SGC7901细胞株．细胞生长和迁移能力显著低于空质粒组（P〈0．01）。结论：成功建立了稳定转染15-PGDH基因的SGC7901细胞株．恢复15-PGDH表达可显著抑制人胃癌细胞的生长和迁移。15-PGDH表达减低或缺失与胃癌的发生、发展密切相关。Background： Our prior studies indicated that the expression of NAD ＋-dependent 15-hydroxyprostaglandin dehydrogenase （15-PGDH） in gastric cancer was relatively low, and transient transfection of 15-PGDH could inhibit the growth and migration of human gastric cancer cells. Aims： To construct human gastric cancer cell line SGC7901 with stable transfection of 15-PGDH gene, and to explore the effect of restoration of 15-PGDH expression on growth and migration of gastric cancer ceils, as well as the relationship between 15-PGDH and the generation and development of gastric cancer. Methods： Eukaryotic expression plasmid peDNA3/15-PGDH was identified by enzyme digestion and sequence analysis, and then transfected into SGC7901 cells by Lipofectamine 2000. Positive clones were selected by G418 and identified with real time polymerase chain reaction （PCR） and Western blotting. The proliferation and migration activities of SGC7901 cell lines with stable transfection of 15-PGDH gene were determined by methyl thiazolyl tetrazolium （MTT） assay and cell scratch assay, respectively. Results： Four SGC7901 cell lines which expressed 15-PGDH stably at high level were obtained by 4-week G418 selection and real time PCR and Western blotting identification. Proliferation and migration capabilities of these four cell lines were significantly inhibited when compared with SGC7901 cells of blank plasmid control group （P〈0.01）. Conclusions： SGC7901 cell lines stably transfected with 15-PGDH gene was successfully constructed. Restoration of 15-PGDH expression can significantly inhibit the growth and migration of human gastric cancer cells. Down-regulation and absence of 15-PGDH are correlated with the generation and development of gastric cancer.

关 键 词：胃肿瘤 羟前列腺素脱氢酶类 转染 细胞增殖 细胞迁移 

分 类 号：R979.1[医药卫生—药品] R335[医药卫生—药学]