EphA2对神经胶质瘤细胞系U251的凋亡﹑增殖﹑迁移和侵袭的研究  

Function Study of EphA2 Gene in The Cell Apoptosis, Growth, Migration and Invasion of Glioma Cell Line U251

作　　者：柏燕燕[1] 史毅[2] 惠国桢[3] 张艳荣[1] 董万利[1] 胡锦[4] 金由辛[2] 

机构地区：[1]苏州大学附属第一人民医院神经内科,苏州215006 [2]中国科学院上海生物化学与细胞生物学研究所,上海200031 [3]苏州大学附属第一人民医院神经外科,苏州215006 [4]上海交通大学附属第六人民医院神经外科,上海200240

出　　处：《生物化学与生物物理进展》2009年第4期464-470,共7页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目(30770824)~~

摘　　要：为了研究EphA2对神经胶质瘤细胞系U251在增殖、凋亡、迁移和侵袭方面所起的作用,用RT-PCR方法检测正常脑组织标本与两种恶性胶质瘤细胞系中EphA2 mRNA表达水平,然后用化学合成的针对EphA2基因的小干扰RNA(siRNA)下调该基因的表达,以检测其在U251中的生物学功能.证实了EphA2基因在正常脑组织标本中的表达水平远低于两种恶性胶质瘤细胞系.把体外化学合成针对EphA2基因的小干扰RNA(siRNA-EphA2)转染入U251细胞后,Western blot,实时定量RT-PCR检测到U251细胞中EphA2蛋白及mRNA表达水平都明显降低,并且细胞增殖受到显著抑制,同时出现了明显的细胞凋亡.伤口愈合实验(检测细胞迁移能力),Transwell小室实验(检测细胞侵袭能力)均表明,下调EphA2的表达后,细胞的迁移和侵袭能力较阴性对照组显著减弱.上述结果表明,在神经胶质瘤U251细胞中,EphA2与其恶性增殖及高度侵染性相关,可作为分子治疗的有效靶点.In order to explore the function of EphA2 gene on the proliferation, apoptosis, motility and invasion of glioma cell line U251. The EphA2 mRNA levels of normal brain and two glioma cell lines U251 and U87 were detected by reverse transcription PCR （RT-PCR）. Then, the function of EphA2 in glioma cell line U251 was analyzed by RNAi technology. The expression level of EphA2 gene was higher in U251 and U87 cells than that in normal brain, siRNA targeting EphA2 was designed and synthesized and then the siRNA was transfected into U251 cell with a high transfection efficiency of 70.31%. 50 nmol/L of siRNA could lead to over 70.0% reduction in EphA2 protein and mRNA level detected by Western blot and RT-PCR respectively. The cell proliferation was notably attenuated and the apoptosis was significantly increased after EhpA2 was downregulated. The migration and invasion of glioma cells were also attenuated when EhpA2 was knocked down by siRNA. EphA2 plays an important role in the malignance of glioma U251 cells and may be a novel molecular marker for the malignance of the glioma cell line U251. EphA2 could be a potential target in gene therapy for glioma.

关 键 词：神经胶质瘤 U251细胞 小干扰RNA EPHA2 

分 类 号：Q7[生物学—分子生物学] R6[医药卫生—外科学]