蜕皮激素诱导MIIP基因的表达对肝癌细胞增殖迁移的影响  

Effect of Ponasterone A induced MIIP Gene Expression on the Proliferation and Migration of Liver Cancer Cells

作　　者：张梅[1] 茹懿[1] 王秦豪[1] 颜广[1] 张耀[1] 李霞[1] 

机构地区：[1]第四军医大学生物化学与分子生物学教研室,陕西西安710032

出　　处：《现代生物医学进展》2017年第9期1610-1614,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(31470803;81572504);肿瘤生物学国家重点实验室(第四军医大学)自主课题(CBSKL2014Z09)

摘　　要：目的:探讨蜕皮激素诱导迁移侵袭抑制蛋白基因(migration and invasion inhibitory protein,MIIP)基因的表达对肝癌细胞SK-Hep-1增殖和迁移能力的影响。方法:采用PCR扩增MIIP基因,连入T载体测序正确后,插入到蜕皮激素可诱导真核表达载体pIND中。将pIND-MIIP和含蜕皮激素受体基因的表达载体pVgRXR以脂质体转染法转染到SK-Hep-1中,经G418和Zeocin双抗生素筛选获得稳定转染细胞株。蜕皮激素Ponasterone A诱导后,采用Western blot检测MIIP表达,CCK-8实验检测细胞增殖,划痕实验检测细胞迁移。结果:蜕皮激素诱导MIIP表达上调后,SK-Hep-1细胞的增殖能力显著下降(P<0.05),细胞的迁移能力明显减弱。结论:建立了MIIP基因可诱导表达系统,证实在肝癌细胞SK-Hep-1中MIIP过表达可抑制细胞的增殖及迁移能力。Objective： To investigate the effect ofPonasterone A induced migration invasion inhibitory protein （MIIP） expression on the proliferation and migration of human liver cancer cell line SK-Hep-1. Methods： MIIP gene was amplified by routine PCR, cloned into pMD18-T vector and confirmed by DNA sequencing, then inserted into the ecdysone-inducible eukaryotic expression plND. SK-Hep-1 cells were co-transfected with pIND-MIIP and pVgRXR, an expression vector contained ecdysone receptor gene, and screened with G418 and Zeocin to obtain the stable clones. Upon treatment with Ponasterone A, one kind of ecdysone, MIIP expression, cell proliferation and migration were analyzed by Western blot, CCK8 and wound healing assay. Results： After Ponasterone A-induced expression of MIIP in SK-Hep-1, cell proliferation was significantly inhibited（P〈0.05）, and cell migration was markedly attenuated. Conclusion： MIIP gene inducible expression system was established and MIIP could inhibit the proliferation and migration of SK-Hep-1 cells.

关 键 词：肝癌 迁移侵袭抑制蛋白 诱导基因表达 迁移侵袭 

分 类 号：R-33[医药卫生] R735.7