碱性成纤维细胞生长因子小干扰RNA与人类免疫缺陷病毒1型病毒蛋白R基因对裸鼠胶质瘤的抑瘤作用  

Anti-glioma effect of combination of bFGF.siRNA and Vpr in nude mice

作　　者：冯学泉[1] 王金环[1] 徐新女[1] 张飚[1] 王淑杰[1] 刘宏胜[1] 林娜[1] 

机构地区：[1]天津市第一中心医院神经外科,300192

出　　处：《中华肿瘤杂志》2010年第10期725-728,共4页Chinese Journal of Oncology

基　　金：基金项目：国家自然科学基金（30672158）

摘　　要：目的 探讨重组腺病毒介导的碱性成纤维细胞生长因子小干扰RNA（bFGF-siRNA）与人类免疫缺陷病毒1型（HIV-1）病毒蛋白R（Vpr）基因联合治疗脑胶质瘤的效果.方法 以人脑胶质瘤LN229细胞感染BALB/c-nu裸鼠,建立动物模型.将30只裸鼠随机分为阴性对照组、空载体组、bFGF-siRNA组、Vpr组和联合治疗组,每组6只,分别定期注射磷酸盐缓冲液（PBS）、空载体腺病毒（rAd5-null）、重组腺病毒rAd5-bFGF-siRNA、重组腺病毒rAd5-Vpr和重组腺病毒rAd5-bFGF-siRNA＋rAd5-Vpr,每隔3 d测量肿瘤体积.治疗后4周处死裸鼠,行HE染色、免疫组化染色、脱氧核糖核苷酸末端转移酶介导的缺口末端标记（TUNEL）法及电镜检查.结果 bFGF-siRNA组、Vpr组和联合治疗组的肿瘤生长抑制率分别为36.9%、37.2%和58.6%,联合治疗组抑瘤效果最显著（P＜0.05）.HE染色、免疫组化染色和凋亡检测均显示,联合治疗组抑制细胞增殖与促进细胞凋亡效果最明显（P＜0.05）.电镜检查显示,联合治疗组肿瘤超微结构的变化最明显.结论 bFGF-siRNA与Vpr联合的基因治疗可以产生明显的协同作用,抑瘤作用较单一治疗明显增强.Objective To study the anti-glioma effect of recombinant adenovirus mediated combined gene therapy of bFGF-siRNA and HIV1-Vpr in vivo. Methods Mouse glioma model was established by injecting 5 × 106 LN229 cells into BALB/c-nu nude mice. 30 nude mice were randomly divided into 5groups： the negative control group, mock group, bFGF-siRNA group, Vpr group and combined therapy group, which at regular intervals were injected with PBS, rAd5-null, rAd5-bFGF-siRNA, rAd5-Vpr, rAd5-bFGF-siRNA plus rAd5-Vpr, respectively. The tumor volume was recorded every third day to draw a growth curve. After four weeks treatment, the mice were killed and specimens were taken. HE,immunohistochemical and TUNEL staining were performed to observe the cell morphology, detect the changes of relevant target proteins and cell apoptosis, respectively. Also the ultrastructural changes were observed by electron microscopy. Results The tumor growth inhibition rates were 36.9%, 37.2% and 58.6% in the bFGF-siRNA group, Vpr group and combined therapy group, respectively, and the combined therapy group showed the most significant effect （P 〈 0.05 ）. Also the results of HE, immunohistochemical and TUNEL staining revealed that the combined therapy group had the best effects on proliferation inhibition and apoptosis induced in glioma cells （ P 〈 0.05 ）. The most significant ultrastructural changes were observed in the combined therapy group. Conclusion The combined gene therapy of bFGF-siRNA with Vpr shows a prominent and synergistic anti-glioma effect compared with that of mono-gene therapy in nude mice.

关 键 词：胶质瘤 小鼠 裸 碱性成纤维细胞生长因子 病毒蛋白R 基因疗法 

分 类 号：R686[医药卫生—骨科学]