RNA干涉CD44基因抑制脑胶质瘤细胞U251的增殖与侵袭性研究  

Knockdown of CD44 by RNAi inhibits proliferation and invasion of glioma cell line U251

作　　者：董超[1,2] 张剑宁[1,2] 杨安钢[3] 任君琳[3] 张雷鸣[1,2] 方丹东[1,2] 陈晓燕[2] 杨韬[3] 郭张燕[3] 

机构地区：[1]海军总医院神经外科,北京100037 [2]第四军医大学西京医院神经外科,陕西西安710032 [3]第四军医大学基础部免疫教研室,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2014年第2期128-132,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(30872978)

摘　　要：目的通过合成靶向分化抗原簇44(CD44)基因的siRNA片段,下调CD44在脑胶质瘤细胞U251中的表达,揭示其在胶质瘤增殖、迁移和侵袭中所起的作用。方法根据CD44基因序列设计并合成的siRNA片段转染U251细胞,利用荧光实时定量(qRT-PCR)检测细胞中CD44基因mRNA水平的变化;蛋白印迹实验(Western blot)检测CD44基因蛋白水平的变化;四甲基偶氮唑蓝染色(methyhhiazolyl tetrazolium,MTT)法检测U251细胞增殖;单细胞划痕愈合实验、Transwell小室侵袭实验检测U251细胞的迁移与侵袭能力变化。结果体外合成的CD44-siRNA能有效抑制U251细胞中CD44基因在mRNA水平与蛋白水平的表达(P<0.05),并抑制细胞的增殖(P<0.05)和迁移侵袭能力(P<0.05)。结论 CD44-siRNA能够有效抑制U251脑胶质瘤细胞的增殖及其迁移侵袭力,为以CD44为靶点的肿瘤基因治疗奠定了基础。Objective The study aims to probe into the effect of cluster of differentiation 44（CD44） gene RNA interference （RNAi） on the proliferation,migration,and invasion of the human glioma cells U251.Methods Human glioma cells （U251） were transfected with the specific siRNA fragment,which was designed to target CD44 mRNA.The expressions of CD44 mRNA and its protein among the transfected cells and the untransfected ones were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot,respectively.The cell growth was examined by methyl thiazolyl tetrazlium （MTT） assay.The changes of cell migration and invasion were observed by monolayer wound healing assay and transwell invasion assay.Results CD44-siRNA inhibited the expression of CD44 gene in mRNA and protein levels of the glioma cells U251 （P 〈0.05）,and inhibited the cell proliferation,migration and invasion of U251 cells （P 〈 0.05）.Conclusion CD44-siRNA can inhibit the proliferation,migration and invasion of U251 glioma cells,so CD44 may be a target for glioma gene therapy.

关 键 词：CD44 RNA干涉 U251 

分 类 号：R735.2[医药卫生—肿瘤]