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背景:反转录病毒载体介导细胞因子杀伤恶性肿瘤细胞为目前肿瘤科研领域的热点,逆转录病毒介导γ-干扰素是否抑制大鼠脑胶质瘤细胞的生长及延长其生存期的作用尚无肯定性结论。目的:观察反转录病毒载体介导的γ-干扰素对大鼠脑胶质瘤细胞的生长抑制作用,探讨与生存期的关系。设计:以动物为研究对象,完全随机设计。单位:一所大学附属医院的神经外科。材料:实验于2002-07/2004-07在哈尔滨医科大学神经外科研究所完成,选择哈尔滨医科大学附属第一临床医学院动物实验中心提供的60只Wistar雄性大鼠。随机分为C6对照组、C6/PA317neo对照组及C6/PA317γ-干扰素治疗组,每组20只。干预:应用反转录病毒载体将鼠γ-干扰素基因转入反转录病毒包装细胞PA317,通过G418抗性筛选,得到高滴度产毒PA317γ-干扰素细胞株。3组大鼠右侧基底核区接种1×106C6细胞。C6对照组:接种后不做其他处理;C6/PA317neo对照组:接种4d后原位注射1×106PA317neo细胞;C6/PA317γ-干扰素治疗组:接种4d后原位注射1×106PA317γ-干扰素细胞。实验观察:①各组大鼠生存期≤60d的情况。②每组动物C6细胞接种后11d及20d后随机取5只处死,取出整个脑组织制备冰冻组织切片,采用SABC免疫组化方法检测肿瘤组织CD4和CD8T淋巴细胞浸润情况。③显微镜下借助图?
BACKGROUND: Retrovirus vector-mediated cytokine killing of malignant tumor cells is currently a hot area in cancer research. Whether retrovirus-mediated interferon-γ inhibits the growth and prolongs the survival of rat glioma cells There is no conclusive conclusion. OBJECTIVE: To observe the inhibitory effect of retroviral vector-mediated interferon-γ on the growth of rat glioma cells and to explore the relationship with the survival time. Design: animal as the research object, completely random design. Unit: Neurosurgery at a university affiliated hospital. MATERIALS: Experiments were performed at Institute of Neurosurgery, Harbin Medical University from July 2002 to July 2004. Sixty Wistar male rats were selected from the First Affiliated Hospital of Harbin Medical University Animal Experimental Center. Randomly divided into C6 control group, C6 / PA317neo control group and C6 / PA317γ-interferon treatment group, 20 rats in each group. Intervention: Transfection of murine interferon-γ gene into retroviral packaging cell line PA317 by retroviral vector and high titer production of PA317γ-interferon cell line by G418 resistance screening. The right basal ganglia of 3 rats were inoculated with 1 × 106C6 cells. C6 control group: no treatment after inoculation; C6 / PA317neo control group: 1 × 106PA317neo cells were in situ injected 4d after inoculation; C6 / PA317γ-interferon treatment group: 4h after inoculation in situ injection of 1 × 106PA317γ-interferon cells . Experimental observation: ① The survival of rats in each group ≤ 60d. ② The C6 cells of each group were sacrificed on the 11d and the 20d after the inoculation, and the whole brain tissues were removed to prepare frozen tissue sections. The infiltration of CD4 and CD8 T lymphocytes in the tumor tissues was detected by SABC immunohistochemistry. ③ under the microscope with the map?