重组质粒pEGFP-N1-EMMPRIN对胃癌SGC-7901细胞侵袭和迁移的影响
Effect of recombinant plasmid pEGFP-N1-EMMPRIN on invasion and migration of gastric cancer SGC-7901 cells
Online published: 2013-03-07
目的 构建人细胞外金属蛋白酶诱导因子(EMMPRIN)重组质粒,观察其在胃癌SGC-7901细胞中的表达及对细胞侵袭和迁移的影响。方法 从结肠癌细胞株SW-480中提取总RNA,通过RT-PCR获得EMMPRIN基因,连接至载体pEGFP-N1,构建重组质粒pEGFP-N1-EMMPRIN,通过脂质体Lipofectamine 2000转染胃癌SGC-7901细胞;RT-PCR和Western blotting分别检测EMMPRIN mRNA和蛋白在SGC-7901细胞中的表达,Transwell实验检测EMMPRIN对SGC-7901细胞侵袭和迁移的影响。结果 重组质粒pEGFP-N1-EMMPRIN构建成功,并在SGC-7901细胞中表达,RT-PCR和Western blotting检测示转染后SGC-7901细胞中EMMPRIN mRNA和蛋白表达均增加,Transwell实验检测示转染后SGC-7901细胞的侵袭和迁移能力均显著增强。结论 EMMPRIN可显著增强肿瘤细胞的侵袭与迁移能力。
关键词: 重组质粒; 细胞外金属蛋白酶诱导因子; SGC-7901; 侵袭; 迁移
亢渝俊, 王 川, 姜 政, 等 . 重组质粒pEGFP-N1-EMMPRIN对胃癌SGC-7901细胞侵袭和迁移的影响[J]. 上海交通大学学报(医学版), 2013 , 33(2) : 155 . DOI: 10.3969/j.issn.1674-8115.2013.02.006
Objective To construct the recombinant plasmid encoding human extracellular matrix metalloproteinase inducer (EMMPRIN), and investigate its expression and effect on invasion and migration of gastric cancer SGC-7901 cells. Methods Total mRNA was extracted from human colon cancer SW-480 cells, EMMPRIN gene was obtained by RT-PCR and cloned into pEGFP-N1 vector, then the recombinant plasmid pEGFP-N1-EMMPRIN was constructed and transfected into human gastric cancer SGC-7901 cells by Lipofectamine 2000. The expression of EMMPRIN mRNA and protein in SGC-7901 cells was detected by RT-PCR and Western blotting respectively, and the effect of EMMPRIN on invasion and migration of SGC-7901 cells was determined with Transewell assay. Results The recombinant plasmid pEGFP-N1-EMMPRIN was constructed and expressed in SGC-7901 cells. RT-PCR and Western blotting indicated that the expression of EMMPRIN mRNA and protein in SGC-7901 cells increased after transfection, and Transwell assay revealed that the invasion and migration of SGC-7901 cells were significantly enhanced after transfection. Conclusion EMMPRIN can significantly enhance the invasion and migration of tumor cells.
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