上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (1): 12-.doi: 10.3969/j.issn.1674-8115.2013.01.003

• 论著（基础研究） • 上一篇    下一篇

EMMPRIN短发夹RNA对SGC-7901细胞侵袭与迁移的影响

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科, 重庆 400016
  • 出版日期:2013-01-28 发布日期:2013-02-06
  • 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。

Influence of short hairpin RNA of EMMPRIN on invasion and migration of 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:2013-01-28 Published:2013-02-06

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摘要：

目的 构建人细胞外金属蛋白酶诱导因子（EMMPRIN）短发夹RNA（shRNA），研究其对人胃癌细胞SGC-7901侵袭与迁移的影响。方法 体外设计合成针对人EMMPRIN的4组小干扰RNA (siRNA)，退火形成双链shRNA，并插入pGenesil-1质粒中。经测序鉴定及RT-PCR和Western blotting筛选出沉默效果明显的干扰质粒，通过脂质体Lipofectamine 2000转染SGC-7901细胞，Transewell法检测重组干扰质粒对SGC-7901细胞侵袭及迁移能力的影响。结果 成功构建并筛选出pshRNA-EMMPRIN干扰质粒。重组干扰质粒转染SGC-7901细胞后，细胞EMMPRIN mRNA和蛋白表达显著降低。Transewell法检测发现，在转染重组干扰质粒的SGC-7901细胞中，侵袭细胞和迁移细胞的个数明显减少。结论 下调EMMPRIN表达能够显著减弱肿瘤的侵袭和迁移能力，为进一步研究EMMPRIN对消化道肿瘤的基因治疗奠定了基础。

关键词: 细胞外金属蛋白酶诱导因子, 短发夹RNA, SGC-7901, 侵袭, 迁移

Abstract:

Objective To construct the short hairpin RNA (shRNA) of extracellular matrix metalloproteinase inducer (EMMPRIN), and investigate its influence on invasion and migration of human gastric cancer SGC-7901 cells. Methods Four groups of human EMMPRIN siRNA were designed and synthesized, double-stranded shRNA was annealed, which was then inserted into pGenesil-1 plasmid. The interference plasmid of EMMPRIN with significant silencing effect was screened by sequencing, RT-PCR and Western blotting, and was transfected into SGC-7901 cells by Lipofectamine 2000. Transwell assay was employed to determine the effect of recombinant interference plasmid on invasion and migration of SGC-7901 cells. Results The interference plasmid pshRNA-EMMPRIN was constructed and screened. After transfection, the expression of EMMPRIN mRNA and protein in SGC-7901 cells significantly decreased. Transwell assay revealed that the number of cells with invasion and migration significantly decreased in SGC-7901 cells after transfection with recombinant interference plasmid. Conclusion The downregulation of EMMPRIN expression can significantly decrease the invasion and migration of tumor cells, which lays a foundation for the further study of gene therapy of tumor with EMMPRIN.

Key words: extracellular matrix metalloproteinase inducer, short hairpin RNA, SGC-7901, invasion, migration