逆转录病毒介导癌基因疗法体外安全实验  

Preclinical Safety Study on Retrovirus Mediated Cancer Gene Therapy in vitro

作　　者：解丽华[1,2] 李殿俊 刘旭[1,2] 夏天 刘威[1,2] 曹雪涛 叶胜龙[1,2] 

机构地区：[1]第二军医大学免疫学教研室 [2]上海医科大学肝癌研究所

出　　处：《哈尔滨医科大学学报》1998年第6期413-415,共3页Journal of Harbin Medical University

摘　　要：以逆转录病毒ｐＬＸＳＮ与人ＩＬ２ｃＤＮＡ基因进行重组，包装ＰＡ３１７细胞，建立逆转录病毒包装体系细胞ＰＡ３１７／ｐＬＩＬ２ＳＮ。用病毒上清液转导人淋巴细胞（ＴＩＬ，ＰＢＬ）和３株人腺癌细胞株（ＳＰＣＡ１，ＭＫＮ４５，ＨｅｐＧ２）对转ＩＬ２基因的淋巴细胞（ＴＩＬ／ＩＬ２，ＰＢＬ／ＩＬ２）和转ＩＬ２基因的人癌细胞（ＳＰＣＡ２／ＩＬ２，ＭＫＮ４５／ＩＬ２，ＨｅｐＧ２／ＩＬ２）进行体外安全性实验，即ｅｎｖ基因检测（ＰＣＲ方法），辅助病毒检测（ＮＩＨ３Ｔ３放大实验），支原体污染检查（ＰＣＲ方法），热原反应（动物实验）。以上实验证明，以ｐＬＸＳＮ逆转录病毒介导外源基因的抗肿瘤基因治疗是安全。The preclinical safety study on retrovirus mediated cancer gene therapy,in vitro were carried out.Retroviral vector pLXSN and human IL-2cDNA was recombinanted,transfected into PA317 cell.The retroviral packaging cell line PA317/pLIL-2SN was constructed.The supernatant of the retroviral packaging cell line PA317/pLIL-2SN transducted to human lympholytes (TIL,PBL) and three human adenocarcinoma cell lines (SPCA 1,MKN-45,HepG 2).In vitro safety assay was performed to transgenic lymphocytes(TIL/IL-2,PBL/IL-2) and transgenic human adenocarcinoma cell lines (transgenic lung adenocarcinoma SPC-A/IL-2,gastric adenocarcinoma MKN-45/IL-2,liver adenocarcinoma HepG 2/IL-2),env gene was determined (PCR method),Helper virus was assessed (NIH3T3 amplication experiment),mycoplasma pollution detection (PCR) and pyrogen reaction (animal experiment) was tested.These preclinical date suggested that retroviral vector pLXSN mediated human cancer gene therapy was safe and viable.

关 键 词：逆转录病毒 基因疗法 辅助病毒 安全试验 肿瘤 

分 类 号：R730.51[医药卫生—肿瘤]