miR-27b影响β-catenin/Tcf-4通路调控U87胶质瘤细胞生物学行为  

miR-27b effect β-catenin/Tcf-4 passway and control the biological behavior of U87 cell

作　　者：杜文众[1] 陈灵朝[1] 崔玉琼[1] 朴星银[1] 雷旭辉[1] 刘幸[1] 康春生[2] 江涛[3] 蒋传路[1] 

机构地区：[1]哈尔滨医科大学附属第二医院神经外科,150086 [2]天津医科大学总医院神经外科,300052 [3]首都医科大学附属北京天坛医院胶质瘤治疗中心,100050

出　　处：《中国微侵袭神经外科杂志》2013年第2期79-82,共4页Chinese Journal of Minimally Invasive Neurosurgery

基　　金：国家863课题(编号:2012AA02A508)

摘　　要：目的研究miR-27b在胶质瘤细胞系U87中的表达、功能及作用机制。方法在多个胶质瘤细胞系及胶质瘤标本中检测miR-27b的表达;利用反义miR-27b转染U87,下调miR-27b的表达,应用流式细胞术和MTT法,评价细胞生长、增殖、凋亡及周期变化;采用荧光素酶法及Western印迹法检测miR-27b作用的初步机制及通路蛋白的变化。结果 miR-27b在胶质瘤细胞系及胶质瘤标本中均呈高表达;转染反义miR-27b后,表达降低86%,MTT法显示转染反义miR-27b后细胞生长受到抑制且凋亡细胞增加13%;荧光素酶实验结果显示:miR-27b发挥作用可能与β-catenin/Tcf-4通路相关,Western印迹法显示通路相关蛋白STAT3、c-myc和Cyclin D1在转染反义miR-27b后表达下降。结论 miR-27b可促进人脑胶质瘤细胞生长、增殖,具有原癌基因的作用,其发挥作用可能与β-catenin/Tcf-4通路密切相关。Objective To investigate the expression, function and action mechanism of miR-27b in glioma cell U87. Methods Expressions of miR-2To were detected in glioma samples and glioma cell lines. And then antisense miR-27b was used to transfect U87 and down-regulate the expression of miR-27b. Meanwhile, flow cytometry and MTT assay were used to evaluate cell growth, proliferation, cell cycle change and apoptosis. Western blotting and luciferase assay were used to detect the change of pathway proteins and preliminary mechanism of miR-27b action. Results miR-27b was up-regulated in glioma samples and glioma cells. After transfection with antisense miR-27b, its expression level decreased by 86%. MTT assay revealed cell growth was inhibited, and the apoptotic cells increased by 13% in glioma cells after antisense miR-27b transfection. Furthermore, luciferase assay suggested action of miR-2To may be related to β-catenin/Tcf-4 pathway. In addition, Western blotting showed the pathway-related proteins STAT3, c-myc and Cyclin D1 were decreased by treatment with antisense miR-27b. Conclusion miR-27b could promote growth and proliferation of human glioma cells and act as a proto-oncogene in human gliomas. The mechanism may be closely related to β-catenin/Tcf-4 signaling.

关 键 词：神经胶质瘤 微RNA 增殖 凋亡 

分 类 号：R739.41[医药卫生—肿瘤] R730.264[医药卫生—临床医学]