| 一种更有效的肝癌靶向性条件复制型腺病毒的构建及体外抑瘤作用 |
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| 引用本文： | 傅强,王慧萍,韦芳,李慧明,陈霞芳,王煜非,黄倩.一种更有效的肝癌靶向性条件复制型腺病毒的构建及体外抑瘤作用[J].现代生物医学进展,2009,9(10). |
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| 作者姓名： | 傅强 王慧萍 韦芳 李慧明 陈霞芳 王煜非 黄倩 |
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| 作者单位： | 上海交通大学医学院附属上海市第一人民医院中心实验室; |
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| 基金项目： | 国家重点基础研究发展规划(973计划)，国家自然科学基金杰出青年项目 |
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| 摘 要： | 目的:分别构建以甲胎蛋白(alpha fetoprotein,AFP)300bp启动子和800bp增强子-300bp启动子调控复制的条件复制型腺病毒(CRAd),对两种病毒的务件复制性以及溶瘤作用进行比较,为肝癌靶向性治疗提供更优良的载体.方法:以HepG2基因组DNA为模板,PCR扩增AFP基因启动子(AFPp)和增强子(AFPe),构建表达质粒pAFPp-EGFPluc和pAFPep-EGFPluc,通过检测报告基因EGFP和luciferase的表达鉴定启动子和增强子的活性后,构建穿梭质粒pDC311-AFPp-E1A,pDC311-AFPep-E1A并包装出腺病毒Ad.AFPp-E1A和Ad.AFPep-E1A.利用Western blot、病毒增殖、细胞病变、细胞活力等鉴定并比较两种病毒的复制能力和溶瘤作用.结果:Ad.AFep-E1A和Ad.AFPep-E1A均可在AFP阳性细胞中选择性复制并且具有一定的溶瘤作用,以后者的选择复制性和溶瘤性更为明显.感染病毒Ad.AFPp-E1A后的HepG2、Hep3B细胞的存活率分别为(54.23±7.13)%、(61.18±12.63)%;感染病毒Ad.AFPep-E1A后的HepG2、Hep3B细胞存活率分别为(26.65±5.43)%、(24.49+3.31)%.结论:被截短的800bp增强子片断,在对AFP启动子起到增强作用的同时,可以为腺病毒包装进更多的治疗基因提供空间,从而为肝癌靶向治疗提供更为良好的条件复制型病毒载体.
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| 关 键 词： | 条件复制型腺病毒载体 甲胎蛋白启动子 甲胎蛋白增强子 |
Construction of a More Effective Conditional Replication-Competent Adenoviral Vectors Targeting Liver Cancer and Its Cancerostatic Effect in Vitro |
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| FU Qiang,WANG Hui-ping,WEI Fang,LI Hui-ming,CHEN Xia-fang,WANG Yu-fei,HUANG Qian.Construction of a More Effective Conditional Replication-Competent Adenoviral Vectors Targeting Liver Cancer and Its Cancerostatic Effect in Vitro[J].Progress in Modern Biomedicine,2009,9(10). |
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| Authors: | FU Qiang WANG Hui-ping WEI Fang LI Hui-ming CHEN Xia-fang WANG Yu-fei HUANG Qian |
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| Institution: | FU Qiang,WANG Hui-ping,WEI Fang,LI Hui-ming,CHEN Xia-fang,WANG Yu-fei,HUANG Qian (Cental Laboratory,the First People's Hospital of Shanghai,School of Medicine,Shanghai Jiaotong University,Shanghai 200080,China) |
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| Abstract: | Objective:To construct two conditional replication-competent adenoviral vectors (CRAs) driven by alpha fetoprotein (AFP)300bp promoter and 300bp promoter+AFP 800bp enhancer respectively,and to find a better CRA vector for liver cancer targeting therapy. Methods:The minimal essential DNA fragment of the AFP gene promoter and enhancer was amplified through PCR from the genome of HepG2 cells. Construct the plasmid pAFPp-EGFPluc and pAFPep-EGFPluc to evaluate the activity of the AFP promoter and enhancer by the... |
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| Keywords: | Replication-competent adenoviral vectors AFP promoter AFPenhancer |
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