研究论文

慢病毒介导的HPV16-E7基因RNAi细胞模型的建立

  • 阿比达·阿不都卡德尔;阿仙姑·哈斯木;赛米热·艾尼瓦尔;阿布力孜·阿布杜拉;盛磊
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  • 1. 新疆医科大学第一附属医院, 乌鲁木齐 830011;2. 新疆医科大学基础医学院, 乌鲁木齐 830011;3. 新疆医科大学新疆地方病分子生物学重点实验室, 乌鲁木齐 830011

收稿日期: 2013-06-28

  修回日期: 2013-11-05

  网络出版日期: 2013-12-18

Construction of Lentivirus-mediated HPV16-E7 Gene RNAi Cell Model

  • ABUDUKADEER Abida;HASIMU Axiangu;AINIWAER Saimire;ABULIZI Abudula;SHENG Lei
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  • 1. The First Afiliated Hospital, Xinjiang Medical University, Urumqi 830011, China;2. College of Basic Medicine, Xinjiang Medical University, Urumqi 830011, China;3. Xinjiang Key Laboratory of Molecular Biology and Endemic Diseases, Xinjiang Medical University, Urumqi 830011, China

Received date: 2013-06-28

  Revised date: 2013-11-05

  Online published: 2013-12-18

摘要

建立HPV16-E7基因的RNAi细胞模型,可为进一步研究HPV16-E7蛋白作用及致癌机制提供物质前提。选择HPV16-E7基因特异的siRNA片段,构建慢病毒siRNA重组表达载体,经293FT病毒包装细胞转染产生重组病毒,并以此感染HPV16阳性SiHa宫颈癌细胞、抗菌素筛选和分子生物学鉴定,建立了稳定表达HPV16-E7-siRNA的RNAi细胞模型。该细胞模型,对目标基因(E7)表达的抑制效率,也以蛋白免疫印迹和RT-PCR确证。由此得出,利用慢病毒载体和HPV16-E7基因特异的siRNA片段,可以建立一种稳定、有效和可行的RNAi细胞模型,为进一步研究E7蛋白的作用及致癌机制奠定了重要的物质基础。

本文引用格式

阿比达·阿不都卡德尔;阿仙姑·哈斯木;赛米热·艾尼瓦尔;阿布力孜·阿布杜拉;盛磊 . 慢病毒介导的HPV16-E7基因RNAi细胞模型的建立[J]. 科技导报, 2013 , 31(35) : 60 -63 . DOI: 10.3981/j.issn.1000-7857.2013.35.011

Abstract

The HPV16-E7 ggene RNAi cell model was constructed. A recombinant lentiviral siRNA expression vector was constructed using a HPV16-E7 oncogene specific siRNA fragment, and an RNAi cell model stably expressing the HPV16-E7-siRNA was established by transfection of 293FT virus-packiging cells with the viral vector followed by infection of HPV16-positive SiHa carcinoma cells with recombinant virus, antibiotica selection and molecular biological characterization. RNAi cell model was successfully established, stably transcribing the HPV16-E7-siRNA fragment, and the inhitory effect on target gene (E7) was also characterized by western blotting and RT-PCR. The RNAi cell model expressing the HPV16-E7 gene-specific siRNA fragment using lentiviral vector was stable, effective and feasible, which lays the foundation for the role study and carcinogenic mechanism of E7 protein.
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