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肿瘤特异性表达早期复制基因E1a腺病毒构建

发表时间:2009-06-15  浏览量:1467  下载量:719
全部作者: 刘珊玲,肖雪,陈新莲,王和
作者单位: 四川大学华西第二医院
摘 要: 目的:重组腺病毒是肿瘤基因治疗中的重要载体,目前用于携带目的基因的腺病毒一般为缺乏早期转录基因E1的复制缺陷型腺病毒,不能有效地将目的基因转入所有肿瘤细胞。端粒酶在肿瘤细胞中有很高活性,而在绝大多数正常细胞中无活性、活性极低或短暂表达,如果能构建包含肿瘤特异性表达的早期复制基因E1a的腺病毒,可解决上述问题。方法:克隆了端粒酶启动子及E1a基因,将其共同装载于穿梭质粒中,采用AdEasy系统,构建了一种用端粒酶启动子控制腺病毒早期转录基因E1a表达的腺病毒,使其在肿瘤细胞中特异性表达E1a.结果:成功构建了用端粒酶启动子控制腺病毒早期转录基因E1a表达的腺病毒。结论:肿瘤中可复制特异性E1a腺病毒的构建,为进一步使带目的基因的腺病毒选择性地在肿瘤细胞中复制的基因治疗策略打下基础。
关 键 词: 肿瘤学;重组腺病毒;端粒酶启动子;E1a基因;基因治疗
Title: Generation of recombinant adenovirus carrying tumor specific expression E1a gene
Author: LIU Shanling, XIAO Xue, CHEN Xinlian, WANG He
Organization: West China Second University Hospital, Sichuan University
Abstract: Objective: Replication-deficient adenovirus, one of the most commonly used vectors in cancer gene therapy, has its advantage of safety when carrying target therapeutic gene into tumor cells. On the other hand, usually it can not efficiently deliver the gene into most of the tumor cells due to lack of replication ability. Telomerase activity is detected in immortal cell lines and an extraordinary diverse set of tumor tissue, but is not detected in most of normal tissues. Thus’ it is regarded as an ideal target for treating proliferation related disease, such as cancer. This research is to generate adenovirus which can specifically replicate in tumor cells. Methods: Telomerase promoter and adenovirus early transcript gene E1a were cloned and constructed into a shuttle plasmid in which E1a expression was controlled by telomerase promoter. The recombination of the shuttle plasmid and viral backbone plasmid was performed in bacteria, and adenovirus was generated in packaging cell line 293. Results: An adenovirus encoding E1a gene under the control of telomerase promoter was generated using AdEasy system. Conclusion: The establishing method of tumor specific expression of E1a could be used in further study of cancer gene therapy strategy regarding tumor specific replication of adenovirus carrying target gene.
Key words: oncology; recombinant adenovirus; telomerase promoter; E1a gene; gene therapy
发表期数: 2009年6月第11期
引用格式: 刘珊玲,肖雪,陈新莲,等. 肿瘤特异性表达早期复制基因E1a腺病毒构建[J]. 中国科技论文在线精品论文,2009,2(11):1147-1152.
 
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