西安交通大学学报（医学版）2011，Vol.32Issue(5)：540-544,5.
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
Packaging and identification of recombinant retrovirus dual cancer-specific expressing superantigen of TSST-1 gene
摘要
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-l-linker-CD80TM, and was co-transfected into packaging cell 293T with helper vector pMB-MLV and pMB-G. LaSRT method was employed to examine the titers of retrovirus. CEA-positive human colon carcinoma cell line Lovo and CEA-negative human cervical carcinoma cell line Hela were infected with recombinant retrovirus respectively. RT-PCR and immunohistochemistry were employed to examine the expression of TSST-1. Results The recombinant retrovirus was harvested and the titer was 1 X 106 CFU/mL. The expression of TSST-1 mRNA and protein in infected Lovo cells under hypoxia condition was significantly higher than that under normoxia condition as confirmed by RT-PCR and immunocytochemistry. It was still expressed after passaged 20 of the infected Lovo cells. Infected Hela cells did not express TSST-1 mRNA or protein under either hypoxia condition or normoxia condition. Conclusion The titer of recombinant retrovirus is enough to infect target cells, and could target and stably express TSST-1 in CEA-positive human colon carcinoma cell. All this provides a foundation for proving TSST-1's effect in killing CEA-positive tumor cells in follow-up tests.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Key words
retrovirus/superantigen/hypoxia induction/CEA-positive tumor/immuno-gene therapy分类
医药卫生引用本文复制引用
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):540-544,5.基金项目
国家自然科学基金资助项目(No.30672070,No.30400430) （No.30672070,No.30400430）
陕西省科学技术研究发展计划项目(No.2009K01-70) （No.2009K01-70）
