内皮抑素和血管生成抑素融合基因的裂解病毒对C6胶质瘤的实验研究  

Therapeutic effects of rHSV-mediated endostatin-angiostatin fusion gene in C6 glioma model

作　　者：王建交[1] 杨立庄[2] 刘福生[1] 金贵善[1] 柴奇[1] 彭小忠(点评)[3] 

机构地区：[1]北京市神经外科研究所,首都医科大学附属北京天坛医院,100050 [2]哈尔滨医科大学附属第二医院神经外科 [3]中国医学科学院基础医学研究所 北京协和医学院基础学院 医学分子生物学国家重点实验室

出　　处：《中华神经外科杂志》2008年第1期69-72,共4页Chinese Journal of Neurosurgery

摘　　要：目的探讨重组单纯疱疹病毒介导的内皮抑素和血管生成抑素（Endo-Angio）融合基因对C6胶质瘤的疗效。方法构建含有Endo-Angio融合基因的重组单纯疱疹病毒，四唑蓝比色法（MTT法）体外观察其C6细胞的裂解作用；Western-blot观察Endo—Angio融合蛋白的表达。建立大鼠C6模型，当肿瘤体积≥2mm^3时，治疗组瘤腔内注射10^7pfu（10μl）病毒，对照组注射等剂量生理盐水。MRI动态观察肿瘤体积变化，组织病理行微血管计数，并记录荷瘤鼠生存时间。结果体外实验示：当感染率（MOI）=1000时，99％的C6细胞死亡；头部MRI发现治疗组胶质瘤生长减慢；荷瘤鼠存活期：对照组（20．3±0．45）d，治疗组（38．3±0．96）d，中位生存期延长（P〈0．01）；HE染色见瘤细胞分散，体积较小，无出血坏死；病理染色示：治疗组和对照组新生血管均值分别为（5．3±1．7）／HP和（15．6±2．8）／HP（P〈0．01）。结论重组单纯疱疹病毒介导的Endo-Angio融合基因不仅通过抑制肿瘤的新生血管生成，而且通过裂解肿瘤细胞双重作用治疗颅内胶质瘤。Objective To explore the inhibitory role of recombinant herpes simplex viral （rHSV） mediated endostatin-Angiostatin fusion gene on the angiogenesis and growth in C6 glioma model. Methods Constructed rHSV vector carrying the Endo-Angio fusion gene. MTr assays were performed to testify the influence of rHSV on the growth of C6 cell line. Western blot showed the band of fusion protein expression. C6 glioma models were established by implanting C6 cells in the brain of Wistar rat, and checked by MRI, If the glioma volume was ≥ 2mm^3, then for virus-gene therapy research. RHSV Endo-Angio was injected intratumorally measured tumor size and morphology were also carried out respectively. Results MTr assay suggested that recombinant simple herpes viral vector could disrupt C6 glioma cells. When MOI = 1000 ,more than 99%C6 cells were killed; HE staining showed that the volume of tumor was smaller and C6 cells was sparsely in the therapy group; Intratumoral microvessel density （MVD） assay showed the number of neovascular was individually （5.3 ± 1.7）/HP and（ 15.6 ±2.8）/HP in the therapy group and control group （P 〈0.01 ） ; Dynamic enhanced MRI scanning showed that the tumor growth was markedly suppressed and the survival time was prolonged in the test group compared with that in the control group （ P 〈 0.01 ）. Conclusion Recombinant herpes simplex viral vector carrying Endo-Angio fusion gene has significantly inhibited the growth of C6 glioma in Wistar rat by the way of inhibiting the angiogenesis and disrupting the cells.

关 键 词：内皮抑素 血管生成抑素 血管生成 C6胶质瘤 病毒-基因治疗 

分 类 号：R73[医药卫生—肿瘤]