慢病毒介导的HIF-1α mODD在肿瘤细胞中的高表达  

Lentiviral Vector Mediated High Expression of HIF-1α mODD in Tumor Cells

作　　者：赵玉娇[1] 龙海亭[1] 潘玥[1] 李华[1] 陈俊英[1] 施海晶 马绍辉[1] 孙强明[1] 

机构地区：[1]北京协和医学院/中国医学科学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,昆明650118

出　　处：《医学研究杂志》2011年第12期28-31,共4页Journal of Medical Research

基　　金：国家自然科学基金面上项目(81171946);中国医学科学院医学生物学研究所人才引进项目(IMB2009RC01)

摘　　要：目的在肿瘤细胞中利用重组慢病毒介导缺氧诱导因子-1α突变体(HIF-1α mODD)在正常氧压下高表达,以模拟肿瘤缺氧微环境下肿瘤细胞中HIF-1α的高表达,为研究HIF-1介导的缺氧信号通路及其在肿瘤细胞中的作用提供实验模型。方法将含HIF-1αmODD基因片段的慢病毒表达质粒pWPI GW/HIF-1α mODD,与pVSVG及pSPAX质粒共转染人胚肾293T细胞,包装制备重组慢病毒,重组病毒经过纯化后直接感染GLC、H157和YTMLC等肿瘤细胞,并在正常氧压条件下培养,通过RT-PCR、免疫印迹等方法检测HIF-1α mODD在细胞中的表达水平。结果重组慢病毒介导HIF-1α mODD在GLC、H157和YTMLC细胞内获得高表达。结论成功模拟了肿瘤细胞在缺氧微环境下高表达HIF-1α的情况,为体外研究HIF-1α在肿瘤血管新生及肿瘤生长中的作用提供途径。Objective To induce high expression of HIF -1α mODD in tumor cells mediated by lentiviral vector in normoxia condi- tion, and simulate high expression of HIF -1αin tumor cells under hypoxia microenvironment in vivo. Methods The packing cell line （human embryonic kidney 293T cells） was eotransfected with pWPI GW/HIF -1α mODD expression vector, pVSVG and pSPAX. The recombinant lentivirus was packaged and amplified, followed by purification, infection of GLC, H157 and YTMLC cells. The expression level of HIF -1α mODD gene was analyzed by RT - PCR and Western blotting. Results RT - PCR and Western blotting results demonstrated that HIF -1αmODD was highly expressed in GLC, H157 and YTMLC cells. Conclusion A model of high expression of HIF -1α in tumor cells hypoxia microenvironment was successfully created. It will provide a method to investigate the role of HIF -1αin tumor angiogenesis and growth in vitro.

关 键 词：HIF-1Α 慢病毒 缺氧微环境 

分 类 号：R651.2[医药卫生—外科学]