| 双靶向诱导超抗原TSST-1基因重组逆转录病毒的包装及鉴定 |
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| 引用本文： | 王伟,孙学军,王炜,郑见宝,禄韶英,任海亮,成亮.双靶向诱导超抗原TSST-1基因重组逆转录病毒的包装及鉴定[J].西安交通大学学报(医学版),2011,32(5):540-544. |
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| 作者姓名： | 王伟 孙学军 王炜 郑见宝 禄韶英 任海亮 成亮 |
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| 作者单位： | 西安交通大学医学院第一附属医院普外科,陕西西安,710061 |
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| 基金项目： | 国家自然科学基金，陕西省科学技术研究发展计划项目 |
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| 摘 要： | 目的包装出携5拷贝缺氧反应元件(5 HRE)增强子和癌胚抗原启动子(CEAp)联合调控的超抗原中毒性休克综合征毒素-1(TSST-1)基因的逆转录病毒,并建立稳定表达TSST-1的人结肠癌Lovo细胞株。方法应用已构建好的逆转录病毒载体pLEGFP-N1-5 HRE-CEAp-TSST-1-linker-CD80TM,与辅助质粒pMB-MLV、pMB-G共转染入293T细胞,LaSRT法检测病毒滴度。包装出的逆转录病毒感染CEA阳性的人结肠癌细胞株Lovo和CEA阴性的人宫颈癌细胞株Hela。应用RT-PCR和免疫组化方法鉴定TSST-1的表达。结果获得重组逆转录病毒滴度约为1×106CFU/mL。RT-PCR及免疫组化证实经病毒感染的Lovo细胞在mRNA和蛋白水平均有TSST-1表达,缺氧环境中的mRNA表达量更高,且细胞传代20次后依然有TSST-1 mRNA表达;经病毒感染的Hela细胞在常氧或缺氧状态下均无TSST-1 mRNA及蛋白表达。结论包装出较高滴度重组逆转录病毒,并能靶向性在CEA阳性肿瘤内稳定表达TSST-1,为后续检验TSST-1对肿瘤的杀伤效应奠定了基础。
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| 关 键 词： | 逆转录病毒 超抗原 缺氧诱导 CEA阳性肿瘤 免疫基因治疗 |
Packaging and identification of recombinant retrovirus dual cancer-specific expressing superantigen of TSST-1 gene |
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| WANG Wei,SUN Xue-jun,WANG Wei,ZHENG Jian-bao,LU Shao-ying,REN Hai-liang,CHENG Liang.Packaging and identification of recombinant retrovirus dual cancer-specific expressing superantigen of TSST-1 gene[J].Journal of Xi‘an Jiaotong University:Medical Sciences,2011,32(5):540-544. |
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| Authors: | WANG Wei SUN Xue-jun WANG Wei ZHENG Jian-bao LU Shao-ying REN Hai-liang CHENG Liang |
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| Institution: | WANG Wei,SUN Xue-jun,ZHENG Jian-bao,LU Shao-ying,REN Hai-liang,CHENG Liang(Department of General Surgery,the First Affiliated Hospital,Medical School of Xi'an Jiaotong University,Xi'an 710061,China) |
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| Abstract: | Objective To package a recombinant retrovirus carrying the superantigen of toxic shock syndrome toxin-1(TSST-1) gene,which is regulated synergistically by 5 copies of hypoxia-responsive element(5HRE) and promoter of carcino-embryonic antigen(CEAp) and identify TSST-1's expression in carcino-embryonic antigen(CEA)-positive human colon carcinoma cell line Lovo.Methods The recombinant retrovirus vector was constructed with the transmembrane superantigen gene of 5HRE-CEAp-regulated TSST-1-linker-CD80TM,and was ... |
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| Keywords: | retrovirus superantigen hypoxia induction CEA-positive tumor immuno-gene therapy |
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