| 携带双调控自杀基因的重组腺相关病毒载体的构建及初步应用 |
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| 引用本文： | 李子博,曾赵军,孙开胜.携带双调控自杀基因的重组腺相关病毒载体的构建及初步应用[J].长沙医学院学报,2008(2). |
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| 作者姓名： | 李子博 曾赵军 孙开胜 |
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| 作者单位： | 长沙医学院医学检验系;中南大学生物科学与技术学院分子生物学研究中心; |
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| 基金项目： | 湖南省教育厅基金资助项目(项目编号:07C151) |
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| 摘 要： | 目的:为构建携带受Tet-On以及VEGF启动子调控自杀基因的重组腺相关病毒载体。方法:用PCR方法扩增VEGF启动子,将其插入pAAV/TRE/HSVtk/Tet-On,形成重组载体pAAV/VEGF/ TRE/HSVtk/Tet-On质粒。进行病毒包装后得到了AAV/VEGF/TRE/HSVtk/Tet-On重组腺相关病毒。用重组腺相关病毒感染乳腺癌细胞株MCF-7和正常乳腺HBL-100细胞,用MTT法及RT-PCR检测在Dox诱导下,GCV对rAAV感染的MCF-7细胞和HBL-100细胞的杀伤作用以及HSVtk基因在MCF-7细胞内的表达情况。结果:在rAAV+Dox+GCV组,GCV对rAAV感染的MCF-7细胞的杀伤作用明显高于rAAV+Dox组,rAAV+Dox组以及HBL-100组,并且RT-PCR结果显示经Dox诱导HSVtk表达较明显。结论:成功构建了携带双调控自杀基因的重组腺相关病毒载体,该病毒载体能有效的感染乳腺癌细胞MCF-7,并能联合GCV治疗,抑制肿瘤细胞生长,而且具有靶向性。
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| 关 键 词： | 基因治疗 腺相关病毒 VEGF启动子 HSVtk基因 Tet-On |
The Construction of Recombinant AAV Vector Mediated Doudle Regulatory Suicide Gene and Its Preliminary Application |
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| LI Zibo,Zeng Zhaojun,Sun Kaisheng Changsha Medical College,Changsha. Molecular Biology Research Center,College of Biological Science , Technology,Central South University,Changsha ,Changsha,Hunan,China..The Construction of Recombinant AAV Vector Mediated Doudle Regulatory Suicide Gene and Its Preliminary Application[J].Journal of Changsha Medical College,2008(2). |
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| Authors: | LI Zibo Zeng Zhaojun Sun Kaisheng Changsha Medical College Changsha Molecular Biology Research Center College of Biological Science Technology Central South University Changsha Changsha Hunan China |
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| Institution: | LI Zibo~1,Zeng Zhaojun~2,Sun Kaisheng 1 Changsha Medical College,410219,Changsha. 2 Molecular Biology Research Center,College of Biological Science , Technology,Central South University,Changsha 410078,Changsha,Hunan,China. |
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| Abstract: | Objective To construct the recombinant AAV vector mediated regulatory suicide gene and elucidate value of its antitumor.Methods:VEGF promoter was inserted into theplasmid pAAV/ TRE/HSVtk/Tet-On and the recombinant plasmid pAAV/VEGF/TRE/HSVtk/Tet-On was constructed, and AAV/VEGF/TRE/HSVtk/Tet-On recombinant adeno-associated virus(rAAV)were then harvested. Purified rAAV was used to infect breast cancer cell strain MCF-7 and normal cell strain HBL-100,killing effects of GCV on MCF-7 cells infected with rAAV an... |
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| Keywords: | Gene Therapy Adeno-associated Virus VEGF promoter HSVtk Gene Tet-On |
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