15-羟基前列腺素脱氢酶基因转染对人胃癌细胞生长和迁移的影响  

Effect of transfection of 15-hydroxyprostaglandin dehydrogenase gene on the growth and migration of human gastric cancer cells

作　　者：申玉翠 张庆华[2] 叶赛[2] 杨燕青[2] 王弢[2] 靖大道[1] 

机构地区：[1]上海交通大学附属第一人民医院消化科,上海200080 [2]生物芯片上海国家工程研究中心,上海201203

出　　处：《肿瘤》2009年第6期532-536,共5页Tumor

基　　金：上海市科学技术委员会基金资助项目(编号:06BZ066)

摘　　要：目的:探讨含有NAD+依赖性l5-羟基前列腺素脱氢酶(NAD+-dependent 15-hydroxyprostaglandin dehydrogenase,15-PGDH)的真核表达质粒(pcDNA3/15-PGDH)转染对人胃癌细胞生长和迁移的影响。方法:Real time PCR检测多种胃癌细胞株中15-PGDH的表达情况。双酶切方法鉴定15-PGDH真核表达质粒pcDNA3/15-PGDH后,应用Lipofectamine 2000将其转染入胃癌细胞SGC7901中。MTT法检测质粒转染对胃癌细胞增殖的影响,划痕实验观察质粒转染对胃癌细胞迁移的影响,软琼脂克隆形成实验观察质粒转染对胃癌细胞克隆形成的影响。结果:SGC7901细胞中15-PGDH表达显著偏低。pcDNA3/15-PGDH质粒转染入SGC7901细胞的效率较高,转染后24 h及48 h时pcDNA3/15-PGDH组的15-PGDH mRNA表达量分别是pcDNA3空质粒组的188倍和271倍。pcDNA3/15-PGDH质粒转染后SGC7901细胞增殖、迁移和克隆形成能力均显著低于空质粒组和未转染组(P<0.05)。结论:15-PGDH基因转染可显著抑制人胃癌SGC7901细胞的增殖、迁移和克隆形成能力,推测15-PGDH可能是抑制胃癌发生、发展的重要因素之一。Objective：This study was designed to explore the effects of transfection of eukaryotic expression vector of NAD ＋ -dependent 15-hydroxyprostaglandin dehydrogenase （pcDNA3/15-PGDH） on the growth and migration of human gastric cancer cell lines. Methods：Real-time PCR was used to detect the mRNA expression of 15-PGDH in 8 lines of human gastric cancer cells. The eukaryotic expression plasmid pcDNA3/15-PGDH was transfected into the gastric cancer cell line SGC7901 with Lipofectamine2000 after identification with double enzyme digestion. Then, MTT assay was used to detect the suppressive effect of 15-PGDH on cell proliferation. Cell scratch healing assay was applied to study the effect of 15-PGDH on cell migration. Soft agar colony formation test was used to observe the effect of 15-PGDH on cell clone formation. Results：The expression level of 15-PGDH mRNA was significantly lower in gastric cancer SGC7901 cells. The transfeetion efficiency of pcDNA3/15-PGDH was higher. The mRNA expression of 15-PGDH in pcDNA3/15- PGDH group was 188 times and 271 times higher than those in pcDNA3 group at 24 h and 48 h after transfection of pcDNA3/15-PGDH. The proliferation, migration, and clony formation of SGC7901 ceils transfected with pcDNA3/15-PGDH were significantly lower than those in empty plasmid group and non-transfected group （ P 〈 0.05 ）. Conclusion： Transfection of 15-PGDH gene significantly inhibited the proliferation, migration, and clony formation of the gastric cancer SGC7901 cells. We supposed that 15-PGDH might be one of the important factors in suppressing the initiation and progression of gastric cancer.

关 键 词：胃肿瘤 羟基前列腺素脱氢酶类 转染 细胞增殖 细胞运动 

分 类 号：R735.2[医药卫生—肿瘤]