摘要
为了实现FOXP2基因在MCF-7细胞中稳定低表达,采用基因工程的方法构建pMAGic7.1-shFOXP2慢病毒干扰载体,并将其与辅助质粒共转染HEK293T细胞,包装shFOXP2干扰慢病毒,收取病毒上清加入到MCF-7细胞中,培养48h,荧光定量PCR检测FOXP2的基因mRNA表达水平,Western Blotting检测FOXP2蛋白表达水平.获得了pMAGic7.1-shFOXP2干扰载体,从而成功建立FOXP2基因稳定干扰的MCF-7细胞株,为进一步研究FOXP2在乳腺癌发生发展中的功能提供了研究材料.
In order to establish a MCF-7 cell line with lentivirus-mediated FOXP2 gene silencing and confirm its interference effect, the lentiviral vector (pMAGic7.1-shFOXP2) was constructed by genetic engineering and cotransfected with pVSVG、Δ8.91 into HEK293T cells, which generated the mature lentivirus, viral supernatant was collected and added to MCF-7 cells after 48 h, the mRNA and protein levels of FOXP2 were examined by qPCR and Western blot. Thus, we constructed and identified the lentiviral recombinant vector pMAGic7.1-FOXP2,successfully established a MCF-7 cell line with the interference of FOXP2 expression, as these findings imply a promising strategy for studying FOXP2 functions in breast cancer progression.
作者
谭拥军
郭明月
向勤
李谕
TAN Yongjun;GUO Mingyue;XIANG Qin;LI Yu(College of Biology,Hunan University,Changsha 410082,China)
出处
《湖南大学学报(自然科学版)》
EI
CAS
CSCD
北大核心
2018年第6期128-132,共5页
Journal of Hunan University:Natural Sciences
基金
国家自然科学基金资助项目(81472718)~~