γ-干扰素基因对大鼠脑胶质瘤细胞生长及生存期的影响(英文)

Effects of gamma-interferon gene on brain gliomas and survival of tumor-carrying rats

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:It has become a hot issue to treat malignant tumors with retroviral mediated cytokines.But it remains a controversy whether retroviral mediated γ interferon(IFN γ ) is valid in killing glioma cells and prolonging the survival of tumor carriers.OBJECTIVE:To observe the effect of retrovital mediated γ interferon on the growth of glioma and the survival of tumor carrying rats. DESIGN:Completely ramdomized trial based on animals. SETTING:Neurosurgery department of a hospital affiliated to a university.MATERIALS:The trial was conducted in the Neurosurgery Institute of Harbin Medical Unicersity from July 2002 to July 2004.The subjects were 60 male Wistar rats provided by the Animal Center of the First Affiliated Hospital of Harbin Medical Unicersity. They were randomly assigned into 3 groups:C6 control group,C6/PA317neo control group and C6/PA317IFN γ treatment group, 20 in each group. INTERVENTIONS:Retroviral vector containing murine IFN γ gene was transferred into retroviral package cell PA317.A high titer PA317IFN γ cell line was obtained after G418 resistance screening. In the right basal ganglion area 1× 106 C6 cells was injected in each of the rats.The rats in C6 groups received no more intervention. Those in C6/PA317neo control group received injection of 1× 106 PA317IFN γ cells in the right basal ganglion 4 days after C6 injection.Those in C6/PA317IFN γ treatment group received injection of 1× 106 PA317IFN γ cells in the right basal ganglion 4 days after C6 injection.Observation:① the survival time of rats within 60 days in each group.② Eleven and twenty days after C6 injection five rats were randomly selected from each group to be kill,and the brains were taken out to make frozen sections for SABC immunohistochemical staining.The CD4 and CD8 T lymphocytes infiltration in the tumors were observed.③ The tumor volume was measured with imaging processing software under microscope. MAIN OUTCOME MEASURES:① The survival time of rats in each group;② Tumor's volume;③ The infiltration of CD4 and CD8 lymphocytes. RESULTS:The mean survival time of rats in C6 and C6/PA317neo control groups was(27.10± 0.82) days and (26.60± 0.93) days respectively(P >0.05),while that in treatment group was(49.30± 4.17) days and there were five rats outlived beyond 60 days(P< 0.01 vs the two controls).The tumor volume in the two control groups was of no difference but the tumor volume in treatment was significantly smaller than that in control groups(P< 0.01).The CD4 and CD8 lymphocytes infiltration in treatment group was more than that in control groups(P< 0.01) and there was no difference between the two controls(P >0.05).It was also revealed through immunohistochemical examination that in treatment group there was Brdu positive cells on the 7th day and they disappeared on the 16th day.But there was no Brdu positive cells in the control groups. CONCLUSION:Intratumor injection of PA317IFN γ induces anti tumor immunal reaction in rats. Retroviral mediated IFN γ inhibits tumor growth and prolongs rat survival.