腺病毒载体介导反义Src基因治疗胶质瘤  

Adenovirus-mediated antisense Src gene therapy for glioma

作　　者：王雪峰[1] 赵鑫[1] 孙鹏鹏[1] 苏平 李殿俊[3] 王智[1] 

机构地区：[1]哈尔滨医科大学附属第四医院神经外科,150001 [2]哈尔滨市第五医院神经外科 [3]哈尔滨医科大学免疫学教研室

出　　处：《中华实验外科杂志》2010年第10期1387-1388,共2页Chinese Journal of Experimental Surgery

基　　金：基金项目：黑龙江省卫生厅基金资助项目（2006-239）;黑龙江省政府博士后基金资助项目

摘　　要：目的 探讨腺病毒载体介导的反义Src基因对胶质瘤生长的作用及其机制.方法 应用携带反义Src基因的重组腺病毒体外感染C6胶质瘤细胞,观察其对细胞形态、生长曲线和克隆形成率的影响.建立大鼠胶质瘤动物模型,原位注射重组腺病毒.观察其治疗作用,Western blot检测肿瘤组织Src、Ras、MAPK蛋白的表达,实时逆转录-聚合酶链反应（Real-time RT-PCR）检测肿瘤组织Caspase-3、Caspase-8 mRNA的表达.结果 体外研究表明感染反义Src基因的C6胶质瘤细胞生长受到显著抑制.体内研究表明应用携带反义Src基因的腺病毒原位注射治疗大鼠胶质瘤能够抑制肿瘤生长,减少Src、Ras、MAPK蛋白的表达,增加Caspase-3、Caspase-8 mRNA的表达.结论 腺病毒载体介导的反义Src基因能够抑制大鼠胶质瘤细胞的生长.Src-Ras-MAPK信号通路参与胶质瘤的生长,抑制该信号通路可以增加凋亡通路关键蛋白Caspase-3、Caspase-8的表达.Objective To investigate the therapeutic effects and the mechanisms of adenovirusmediated gene transfer of antisense Src for rat C6 glioma. Methods C6 glioma cells were infected with recombinant adenovirus containing antisense Src gene. Cell morphology, cell growth curve and cloning efficiency assay were examined. The intracranial C6 glioma animal model was established in immunocompetent Wistar rats. Recombinant adenoviruses containing antisense Src gene were stereotactically injected into the tumor areas. Tumor volumes were measured. Western blotting was used to detect Src, Ras and MAPK proteins expression. Caspase-3 and Caspase-8 mRNA expression in tumor was quantified by real-time reverse transcription-polymerase chain reaction （RT-PCR）. Results Cell growth and cloning efficiency of C6 glioma cells infected with recombinant adenovirus containing antisense Src gene were significantly decreased.In situ injecting recombinant adenovirus containing antisense Src gene suppressed tumor growth. In vivo investigation showed Src, Ras and MAPK proteins were significantly decreased in the recombinant adenovirus containing antisense Src gene-treated group, and Caspase-3 and Caspase-8 expression was increased. Conclusion Adenovirus-mediated gene transfer of antisense Src inhibited C6 glioma cells growth in vivo and in vitro. Src-Ras-MAPK signal pathway played an important role in the progression of glioma. MAPK signal pathway blockade could augment apoptosis proteins Caspase-3 and Caspase-8 expression. Our results suggested adenovirus-mediated gene transfer of antisense Src was a promising target for gene therapy of malignant brain tumors.

关 键 词：胶质瘤 基因治疗 反义Src 腺病毒载体 

分 类 号：R73[医药卫生—肿瘤]