上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (2): 155-.doi: 10.3969/j.issn.1674-8115.2013.02.006

• 论著（基础研究） • 上一篇    下一篇

重组质粒pEGFP-N1-EMMPRIN对胃癌SGC-7901细胞侵袭和迁移的影响

亢渝俊, 王 川, 姜 政, 王丕龙   

  1. 重庆医科大学附属第一医院消化内科, 重庆 400016
  • 出版日期:2013-02-28 发布日期:2013-03-07
  • 通讯作者: 姜 政, 电子信箱: jiangz1753@163.com。
  • 作者简介:亢渝俊（1977—）, 女, 主治医师, 硕士生；电子信箱: 330273776@qq.com。

Effect of recombinant plasmid pEGFP-N1-EMMPRIN on invasion and migration of gastric cancer SGC-7901 cells

KANG Yu-jun, WANG Chuan, JIANG Zheng, WANG Pi-long   

  1. Department of Gastroenterology, the First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China
  • Online:2013-02-28 Published:2013-03-07

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摘要：

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

Abstract:

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.

Key words: recombinant plasmid, extracellular matrix metalloproteinase inducer, SGC-7901, invasion, migration