摘要
研究了反转录病毒介导的ＨＳＶ－ｔｋ／ＡＣＶ系统基因治疗脑肿瘤的安全性。首先采用ＰＣＲ、反转录ＰＣＲ（ＲＴ／ＰＣＲ）、Ｓｏｕｔｈｅｒｎ杂交以及补救分析方法，对ＨＳＶ－ｔｋ病毒包装细胞（ＴＫ／ＶＰＣ）、Ｃ６转ＴＫ基因细胞和实验动物血液进行了检测。结果表明，所采用的反转录病毒基因转移系统没有产生野生型病毒，是安全的；另一方面，采用细胞接种、Ｘ－ｇａｌ染色、ｔｋ基因的ＰＣＲ和病理切片观察方法，对ＨＳＶ－ｔｋ／ＡＣＶ系统本身的安全性进行了探讨，结果表明ＨＳＶ－ｔｋ／ＡＣＶ系统用于动物基因治疗，ＡＣＶ及其代谢产物没有发现明显的毒副作用和病理改变。反转录病毒介导的ＨＳＶ－ｔｋ／ＡＣＶ系统是安全的。
The safety assessments on retroviral mediated HSV tk/ACV gene therapy for glioma have been made. First, polymerase chain reaction(PCR), reverse transcripted PCR(RT/PCR), Southern blot and rescue assay were used to detect replication competent retrovirus (RCR) in HSV TK virus producing cell line (TK/VPC), C6TK and SHGTK genetically modified cell and blood samples. The retrovirus mediated gene delivery system has not produced RCR. On the other hand, cell implantation, PCR for HSV tk, X gal staining and histomicrosection were used to deal with safety assessments of HSV tk/ACV system in mice, rats and rabbits. Virus producing cell PA317/BAG, PA317/TK can not live more than 1 month. No inflammatory response and tumor occurred. No visible side effects or pathological changes were attributed to the gene transfer system. This study indicated that HSV tk/ACV system is safe for the treatment of brain tumors.
出处
《高技术通讯》
EI
CAS
CSCD
1998年第8期50-55,共6页
Chinese High Technology Letters
基金
上海市科委启明星资助项目
关键词
脑肿瘤
反转录病毒载体
基因治疗
安全性
Retroviral vector, HSV tk/ACV system, RCR, Gene therapy, Safety
