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慢病毒介导MCF-7细胞中FOXP2基因沉默体系的建立

Construction of FOXP2 Gene Silencing System in MCF-7 Cells by Lentivirus Infection
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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)~~
关键词 FOXP2 慢病毒载体 RNA干扰 乳腺癌MCF-7细胞 the forkhead box p2 lentivirus vector RNA interference breast cancer MCF-7 cells
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