EMMPRIN短发夹RNA对SGC-7901细胞侵袭与迁移的影响  

Influence of short hairpin RNA of EMMPRIN on invasion and migration of SGC-7901 cells

作　　者：亢渝俊[1] 王川[1] 姜政[1] 王丕龙[1] 

机构地区：[1]重庆医科大学附属第一医院消化科,重庆400016

出　　处：《上海交通大学学报（医学版）》2013年第1期12-18,24,共8页Journal of Shanghai Jiao tong University:Medical Science

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。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.

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 

分 类 号：R735.2[医药卫生—肿瘤]