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在腺病毒介导的HSV-tk/NAS系统脑肿瘤基因治疗研究过程中,建立了野生型腺病毒(RCA)的检测系统,主要包括腺病毒E1区的PCR以及野生型病毒的细胞病理效应观察这两种方法,PCR的灵敏度为1个293细胞所含有的E1片段数。对基因治疗脑肿瘤所采用的重组腺病毒AdTK及其感染的细胞进行RCA的检测,没有发现RCA。通过离体实验首次发现,重组腺病毒对脑肿瘤细胞的杀伤作用明显高于正常的脑胶质细胞;大鼠和兔脑内原位注射重组腺病毒AdTK,并腹腔注射GCV,正常脑组织只有轻微的病理改变,脑内可检测的重组腺病毒不超过4周;小鼠和大鼠尾静脉注射重组腺病毒和腹腔GCV治疗,除肺和肝脏有GCV引起的炎症反应外,其他主要脏器没有病理改变。
In the process of adenovirus-mediated HSV-tk / NAS system brain tumor gene therapy, the detection system of wild-type adenovirus (RCA) was established, which mainly included the PCR of adenovirus E1 region and the cytopathological effects of wild-type virus For both methods, the sensitivity of PCR is the number of E1 fragments contained in one 293 cell. Detection of RCA by recombinant adenovirus AdTK and its infected cells used in gene therapy brain tumors showed no RCA. It was first found by in vitro experiments that the killing effect of recombinant adenovirus on brain tumor cells was significantly higher than that of normal glial cells; in situ injection of recombinant adenovirus AdTK and intraperitoneal injection of GCV in rats and rabbits, normal brain tissue only Mild pathological changes, the brain can detect recombinant adenovirus less than 4 weeks; mouse and rat tail vein injection of recombinant adenovirus and abdominal GCV treatment, in addition to lung and liver GCV-induced inflammatory response, the other major organs No pathological changes.