慢病毒介导稳定敲低Smurf1细胞株的构建及对细胞迁移的影响  

Establishment of two human tumor cell lines with lentivirus-mediated stably Smurf1 silencing and their effect on cell migration

作　　者：韦荣飞 郭静[1] 李梦媛[1] 朱瑞敏[1] 杨星九 高苒[1] 

机构地区：[1]中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,北京100021

出　　处：《中国比较医学杂志》2017年第4期46-51,共6页Chinese Journal of Comparative Medicine

基　　金：协和青年基金资助;中央高校基本科研业务费专项资金资助(3332016077);中国医学科学院医学实验动物研究所基本科研业务费专项资助(2016ZX310033)

摘　　要：目的构建慢病毒介导稳定敲低Smurf1的HeLa和A549细胞株并检测敲低Smurf1细胞迁移的影响。方法将包装好的Smurf1敲低慢病毒感染HeLa和A549细胞,7 d后进行嘌呤霉素抗性筛选阳性细胞,Western blot和qPCR检测敲低效果,并进行Transwell检测Smurf1敲低对细胞迁移的影响。结果利用干扰慢病毒系统成功构建稳定敲低Smurf1的HeLa和A549细胞株,稳定敲低Smurf1抑制细胞的迁移速率。结论敲低Smurf1抑制细胞迁移。Objective To establish lentiviral expression vectors for Smurfl silencing and assess the effects of Smurfl silencing on cell migration. Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smnrfl silencing respectively. After 7 days, the stable cell lines with Smurfl silencing were obtained after puromycinresistance screening, enrichment and expansion. The intracellular gene and protein levels of Smurfl were detected by qPCR and western blot. Transwell assay was used to assess the effect of Smurfl silencing on cell migration. Results The stable cell lines with Smurfl silencing are constructed successfully. Silencing of Smurfl down-regulated cell migration rate detected by Transwell assay. Conclusion Smurfl promotes cell migration.

关 键 词：Smurf1 慢病毒 RNA干扰 细胞迁移 

分 类 号：R33[医药卫生—人体生理学]