腺病毒介导的BmK CT基因抑制神经胶质瘤U251细胞迁移及诱导凋亡的研究  

Effects of Ad-BmK CT on Apoptosis and Inhibition of Migration of Glioma Cells U251

作　　者：杜军[1] 付月君[1] 张超[2] 胡涛[3] 梁爱华[1] 

机构地区：[1]山西大学生物技术研究所,山西太原030006 [2]中国科学院生物物理研究所,北京100101 [3]山西省人民医院神经外科,山西太原030006

出　　处：《山西大学学报（自然科学版）》2013年第2期253-260,共8页Journal of Shanxi University(Natural Science Edition)

基　　金：国家自然科学基金(31272100;31071924);国家"863计划"项目(2012AA020809);太原市科技攻关计划(120122);山西省高校优秀青年学术带头人(20120302)

摘　　要：以腺病毒为载体介导东亚钳蝎氯离子通道神经毒素(BmK CT)感染人脑神经胶质瘤细胞株U251,研究其对细胞迁移及凋亡的影响,并探讨可能的作用机制.首先以含BmK CT基因的腺病毒Ad-BmK CT及对应的腺病毒空载体转染U251细胞筛选出最佳感染浓度,随后利用Boyden Chamber实验和划痕实验检测了BmK CT对U251细胞迁移能力的影响,MTT检测了细胞存活能力,用Annexin V-Cy5和PI双标记流式检测分析细胞凋亡情况,并通过Western-blot检测凋亡相关蛋白分析可能的凋亡机制.结果显示,U251细胞感染Ad-BmK CT 48h后,迁移能力受到明显抑制;细胞变圆漂浮,Western-blot检测发现细胞色素c开始释放,caspase-3表达上调.Chlorotoxin （CTX） was reported to inhibit glioma invasion and metastasis. BmK CT, a venom peptide with 68% homology to chlorotoxin,was expressed by using adenovirus vector to study its effects on the migration and apoptosis of human U251 glioma cell. Firstly,Boyden Chamber and wound healing as- says showed that expression bf BmK CT restrained migration in U251 cells. Secondly,cells infected by Ad- BmK CT showed remarkably reduced proliferation rate in MTT assay, which might due to the promotion of apoptosis in BmK CT positive cells. Finally, BmK CT facilitated cytochrome c release and upregulated caspase-3,which might account for the increased apoptosis rates in BmK CT positive cells. This study pro- vided an experimental evidence that Ad-BmK CT might be a potential succedaneum of CTX for the treat- ment of glioma.

关 键 词：东亚钳蝎 氯离子通道神经毒素 腺病毒 神经胶质瘤 

分 类 号：Q78[生物学—分子生物学]