沉默Maspin基因对人乳腺癌细胞MCF-7侵袭迁移能力的影响  

Effect of Maspin Gene Silencing on Invasion and Migration of Human Breast Cancer Cell Line MCF-7

作　　者：李丽萍[1] 刘劲松[1] 

机构地区：[1]湖北省仙桃市第一人民医院肿瘤科,仙桃433000

出　　处：《世界科技研究与发展》2013年第3期422-426,共5页World Sci-Tech R&D

摘　　要：目的探讨靶向沉默maspin基因对乳腺癌MCF-7细胞侵袭和迁移能力的影响。方法构建针对maspin基因的具有荧光蛋白表达的shRNA真核表达载体,稳定转染入乳腺癌细胞MCF-7中。通过划痕实验和Transwell小室侵袭实验分别观察转染前后细胞迁移及侵袭能力的影响。分别用RT-PCR及Western Blot检测转染重组质粒前后细胞maspin mRNA和蛋白表达情况。结果成功构建表达质粒pGenesil-HK、pGenesil-maspin-1和pGenesil-maspin-2,并成功稳定转染MCF-7细胞。与未转染组和pGenesil-HK组相比,转染pGenesil-maspin-1和pGenesil-maspin-2后的MCF-7细胞划痕伤口愈合速度加快(P<0.05),细胞侵袭至小室下层的细胞数明显高于对照组(P<0.05)。RT-PCR及Western Blot结果表明转染pGenesil-maspin-1和pGenesil-maspin-2后的细胞maspin的mRNA和蛋白表达明显下降。结论靶向maspin基因的RNA干扰能够下调maspin基因在乳腺癌MCF-7细胞中的表达,并能增强肿瘤细胞的侵袭和迁移能力。Objective To explore the effects of down-regulating maspin expression by RNA interference on migration and invasion in human breast eaneer MCF-7 cells. Methods The recombinant eukaryotic expression plasmids pGenesil-HK,pGenesil-maspin-1 and pGenesil-maspin- 2 were constructed anti transfected into MCF-7 cells. The migration and invasion ability of MCF-7 ceils was detected by scratch test and Tran- swell chamber assay,respectively. The mRNA and protein expressions of maspin in MCF-7 cells were detected by RT-PCR and Western Blot, respectively. Results We successfully constructed the recombinant eukaryotie expression plasmids pGenesil-HK, pGenesil-maspin-1 anti pGenesil-maspin-2 and transfected into MCF-7 cells. Compared with untransfected group and pGenesil-HK group,the migrating speed of the cells speed up （ P 〈 O. 05 ） ,anti the number of cells passing through the Transwell chamber increased significantly （ P 〈 O. 05 ）. Compared with untranst^cted group and pGenesil-HK group, the mRNA and protein expression of maspin in MCF-7 cells transfected with pGenesil- maspin-1 or pGenesil-maspin-2 were significantly inhibited. The inhibition rate were 41.87% ,68.07% and 45.21% riO. 25% ,respectively. Conclusion RNA interference targeting maspin gene can increased the ability of cell migration and invasion by effectively down-regulated the expression of maspin in MCF-7 ceils.

关 键 词：RNA干扰 MASPIN MCF-7细胞 侵袭 迁移 

分 类 号：R737.9[医药卫生—肿瘤]