摘要
目的 探讨G蛋白耦联受体137（GPR137）对胶质瘤细胞增殖、克隆形成能力和细胞周期的影响.方法 半定量逆转录-聚合酶链反应检测胶质瘤细胞U251、U-87中GPR137 mRNA的表达.构建包含GPR137基因的慢病毒载体,转染人人胶质瘤U251细胞,应用实时定量聚合酶链反应和Western blot法验证GPR137 mRNA和蛋白的表达.应用噻唑蓝比色法、克隆形成实验和流式细胞分析法评价GPR137对U251细胞增殖、克隆形成和周期分布的影响.结果 GPR137在两种胶质瘤细胞中均显著表达[在U251中表达增加了（66.2±1.8）％,在U87中表达增加了（33.8±3.4）％）].GPR137-shRNA对U251的抑制率达到76.5％.与对照组比较,实验组胶质瘤细胞的增殖减少了37.4％;克隆实验表明克隆形成能力明显受到抑制;流式细胞分析显示周期分布为：G0/G1：52.18％,S：41.05％,G2/M：6.77％,周期停滞在S期.结论 GPR137在胶质瘤细胞中高表达,沉默GPR137基因可明显抑制U251细胞的增殖及克隆,并阻滞其细胞周期于S期.
Objective To investigate the effect of G protein-coupled receptor 137 （GPR137） on proliferation,colony and cell cycle of glioma cell line U251.Methods Expression levels of GPR137 mRNA in U251,U-87 glioma cells were detected by semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR） ; we used lentivirus-mediated small interfering RNAs （siRNAs） to knock down GPR137 expression in U251 cells.Expression levels of GPR137 mRNA and protein were determined by Real-time PCR and Western blotting; cell proliferation was examined by methyl thiazol tetrazolium （MTT） assay; the colony-forming capacity was verified by colony formation assay; the cell cycle distribution was detected by flow cytometric analysis.Results We found that GPR137 mRNA was highly expressed in U251 and U-87 cells [increased （66.18 ± 1.80） % and （33.82 ± 3.40） % respectively].After transfection of Lv-sh GPR137 into U251 cells,as compared with control group,inhibition rate of Lv-sh GPR137 reached 76.5%.Inhibition of GPR137 expression in U251 cells by RNAi significantly impaired cell proliferation by 37.4%,the colony-forming capacity,and significantly arrested the cell cycle in S phase （G0/G1：52.18%,S：41.05%,G2/M：6.77%）.Conclusion GPR137 is highly expressed in glioma cell lines.Furthermore,GPR137 play a critical role in cell proliferation,colony formation and cell cycle distribution of U251 cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第9期1886-1888,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81072066）
