微小RNA-140-5p抑制胶质瘤细胞株U251、U87增殖和侵袭性的研究  

Inhibitory effects of microRNA-140-5 p on proliferation and invasion of human glioma cell lines U251 and U87 in vitro

作　　者：吴淳[1] 李炎炎[1] 李学涛[1] 吴至武 程哲[1] 韩勇[1] 孙婷[1] 周幽心[1] 杜子威[1] 

机构地区：[1]苏州大学附属第一医院神经外科脑神经研究室,215006

出　　处：《中华实验外科杂志》2015年第9期2059-2063,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81372689）;江苏省卫生厅重点科研课题资助项目（K201106）

摘　　要：目的 分析微小RNA（miRNA,miR）-140-5p在人脑胶质瘤组织中的表达水平,观察miR-140-5p对胶质瘤细胞株增殖、凋亡与侵袭的影响.方法 通过实时定量聚合酶链反应（Real-time PCR）检测人脑胶质瘤组织与U251、U87MG细胞株中miR-140-5p的表达;用脂质体将miR-140-5pmimics转染至U251与U87MG细胞,同时设立空白及阴性对照组,Real-time PCR检测转染前后miR-140-5p在细胞中的表达水平.应用荧光显微镜观察转染效率,细胞计数试剂盒（CCK-8）法检测细胞增殖能力,流式细胞仪分析细胞周期与凋亡;Transwell实验判断U251、U87MG的侵袭能力.结果 miR-140-5p在人脑胶质瘤组织及胶质瘤细胞株中表达明显低于非瘤脑组织,并且随着肿瘤恶性程度不断增加,miR-140-5p表达逐渐降低（非瘤脑组织、低级别与高级别胶质瘤相对表达量分别为1.013、0.375和0.130,P＜0.01）.U251和U87MG细胞转染miR-140-5p mimics后细胞的增殖与侵袭能力明显受到抑制,其中相对增殖率分别为61.22％与58.44％,穿膜细胞数亦减少了约50％,细胞周期阻滞于G0/G1期（P＜0.01）.转染miR-140-5p mimics后胶质瘤细胞株的凋亡率明显升高＞12％（P＜0.01）.结论 miR-140-5p在胶质瘤组织中低表达,过表达miR-140-5p可以抑制胶质瘤细胞株U251和U87MG的增殖,阻滞细胞周期,促进其细胞凋亡,并可有效抑制其侵袭力.Objective By exploring the expression of microRNA （miR）-140-5p in human glioma tissues,to investigate the effects of miR-140-5p on proliferation apoptosis and invasion of glioma cell lines.Methods By real-time fluorescence quantitative PCR （Real-time PCR） detects the expression ofmiR-140-5p in human glioma tissues and U251,U87MG cell lines.miR-140-5p mimics were transfected into U251 and U87MG cells by Lipofectamine 2000.At the same time,the blank control group and negative control group were established.The expression level of miR-140-5p was identified by Real-time PCR.After transfection,transfection efficiency was observed by fluorescent microscope.The ability of cell proliferation was detected by cell counting kit-8 （CCK-8）;cell cycle and apoptosis were detected by flow cytometry;the cell invasive ability was evaluated by Transwell assay.Results Real-time PCR showed that miR-140-5p expression in non-cancerous brain tissues was lower than in human brain glioma tissues and glioma cell lines,while it decreased with the increasing degree of malignancy in glioma （the relative expression of non-cancerous brain tissues,low grade and high grade glioma were 1.013,0.375 and 1.130,P ＜0.01）.After over-expression of miR-140-5p,the U87MG and U251 cells proliferation and invasion ability were obviously inhibited （P ＜ 0.01）.Among them,the relative proliferation rate were 61.22％ and 58.44％,the cell nember of who cross cell membrane fell by almost half.By flow cytometry on cell cycle analysis,results showed that cell cycle was arrested in G0/G1 phase and S phase was decreased （P ＜ 0.01）.The cell apoptosis was significantly increased larger than 12％ after over-expression of miR-140-5 p （P ＜ 0.01）.Conclusion miR-140-5 p in glioma tissues is low expression,over-expression of miR-140-5p can inhibit glioma cell proliferation,arrest cell cycle,promote its apoptosis,and can effectively inhibit the invasiveness.

关 键 词：微小RNA-140-5p 胶质瘤 细胞增殖 凋亡 侵袭 

分 类 号：R730.43[医药卫生—肿瘤]