MiR-29a下调共刺激分子B7-H3的表达及其对脑胶质瘤细胞侵袭的影响  

MiR-29a inhibited costimulatory molecule B7-H3 expression and the invasion of glioma growth

作　　者：高兵[1,2] 陈翰卿[1,2] 时正鹏[1,2] 孙静[3] 严茹红[4] 傅丰庆[5] 张学光[1,2,5] 

机构地区：[1]苏州大学基础医学与生物科学学院,江苏苏州215000 [2]苏州大学医学生物技术研究所,江苏苏州215000 [3]苏州卫生职业技术学院,江苏苏州215000 [4]苏州大学附属第二医院检验科,江苏苏州215000 [5]苏州大学附属第一医院江苏省临床免疫研究所,江苏苏州21500

出　　处：《中国肿瘤生物治疗杂志》2015年第1期28-33,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.30901313;No.31100626;No.81301494)~~

摘　　要：目的:探讨miR-29a调控共刺激分子B7-H3在脑胶质瘤中的表达及其对脑胶质瘤细胞侵袭能力的影响。方法:通过Real-time PCR检测miR-29a和B7-H3在正常脑组织、脑胶质瘤组织及人胶质瘤细胞株U87中的表达,并利用脂质体将miR-29a的模拟物(mimics)和抑制剂(inhibitors)转入U87细胞,流式术验证miR-29a对B7-H3表达的调节效果;采用CCK-8和Transwell实验观察miR-29a对U87细胞的增殖和侵袭能力的影响,并通过流式术分析miR-29a干预前后U87细胞上与细胞侵袭相关的化学趋化因子的表达变化,以miRtarbase等软件预测miR-29a与CXCR4的结合能力。结果:胶质瘤组织及细胞株中miR-29a低表达而B7-H3 mRNA高表达,且均与瘤组织病理分级相关。转染miR-29a mimics可以有效下调U87细胞中B7-H3 mRNA的表达。转染miR-29a mimics可以显著抑制U87细胞的侵袭能力(P<0.05),但对细胞增殖并无显著影响。miR-29a过表达可同时下调U87细胞中CXCR4的表达,但软件分析CXCR4基因上并不存在miR-29a的结合位点。结论:miR-29a可有效下调B7-H3分子的表达,进而抑制脑胶质瘤细胞的侵袭能力,其作用机制可能与CXCR4途径相关。Objective: To determine the expression and possible roles of miR-29 a and B7-H3 in glioma growth and invasion. Methods: Glioma tissue specimens were collected from 19 patients who underwent surgical glioma resection in the Department of Neurosurgery,Soochow University-Affiliated First Hospital between September,2006 and December,2010.Levels of miR-29 a and B7-H3 mRNAs in the tissue specimens and human glioma U87 cells were determined by Real-time PCR. U87 cells were transfected with miR-29 a mimics,and B7-H3 expression and invasive capacity in the transfectants were assessed by flow cytometry and transwell migration assay,respectively. The expression of invasion-related chemical chemokines was analyzed by flow cytometry,and the ability of miR-29 a to bind to CXCR4 was predicted by the miRtarbase software. Results: In glioma tissue specimens,miR-29 a and B7-H3 mRNA levels were inversely correlated. In vitro,miR-29 a down-regulated B7-H3 mRNAs expression in U87 cells and miR-29a-mediated down-regulation of B7-H3 resulted in a significant decrease in the invasive ability but not proliferative activity in U87 cells. Parallel to down-regulation of B7-H3 expression,CXCR4 expression was also down-regulated in U87 cells transfected with miR-29 a mimics. No miR-29 a binding sites were detected in the CXCR4 gene. Conclusions: In human glioma,miR-29 a can effectively down-regulate B7-H3 expression and inhibit invasive activity. It is likely that these effects of miR-29 a were at least attributable dwonregulated expression of CXCR4.

关 键 词：B7-H3 miR-29a 胶质瘤 U87细胞 侵袭 增殖 CXCR4 

分 类 号：R739.41[医药卫生—肿瘤]