微小RNA-874对U87胶质瘤细胞增殖和凋亡的影响

Effect of microRNA-874 on proliferation and apoptosis in U87 glioma cells

目的 观察微小RNA-874（miR-874）对胶质瘤细胞U87增殖和凋亡的影响.方法 通过实时荧光定量聚合酶链反应（FQ-PCR）检测6例手术获取的非肿瘤脑组织和12例胶质母细胞瘤组织miR-874的表达;体外培养人胶质瘤细胞U87分别转染无义序列（对照组）和合成miR-874模拟体（实验组）,FQ-PCR检测转染后U87细胞miR-874的表达;采用噻唑蓝（MTT）实验评估U87细胞的增殖;采用流式细胞术检测U87细胞周期和凋亡.结果 miR-874在胶质母细胞瘤组织的表达量明显低于非肿瘤脑组织,合成miR-874模拟体可以上调U87细胞miR-874的表达水平（实验组：638.00±80.63;对照组：0.17±0.04,P＜0.05）,进而抑制U87细胞的增殖（48 h：实验组：0.357±0.026;对照组：0.589±0.021,P＜0.05;72 h：实验组：0.522±0.034;对照组：0.834±0.035,P＜0.05）;上调U87细胞miR-874的表达导致G0/G1期细胞比例增加[实验组：（56.28±1.62）％;对照组：（48.57±0.88）％,P＜0.05],同时促进U87细胞凋亡[实验组：（12.50±0.66）％;对照组：（4.91±0.15）％,P＜0.05].结论 miR-874在胶质母细胞瘤中低表达.miR-874与胶质瘤发生、发展密切相关,可能是胶质瘤诊断和治疗的有效靶点.

Objective To observe the effects of microRNA （miR）-874 on proliferation and apoptosis in U87 glioma cells.Methods The expression of miR-874 was detected in 12 cases of gliomas and 6 non-tumor control brain tissues by using real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）.The expression of miRNA-874 was tested by FQ-PCR after synthesized miR-874 mimics was transfected into U87 glioma cells and cell proliferation was detect by methyl thiazol tetrazolium （MTT）.Flow cytometry was applied to detect the cell cycle and apoptosis.Results MiR-874 was down-regulated in glioblastoma samples compared with non-tumor control brain tissues.The expression of miR-874 increased significantly after transfection of miR-874 mimics in U87 cells compared to control group （638.00 ±80.63 vs.0.17 ±0.04,P 〈0.05）,meanwhile suppressed cells proliferation （48 h：0.357 ± 0.026 vs.0.589 ± 0.021,P 〈 0.05 ; 72 h：0.522 ± 0.034 vs.0.834 ± 0.035,P 〈 0.05）.Up-regulation of miR-874 raised the percentage of G0/G1 phase cells [（56.28 ± 1.62）% vs.（48.57 ±0.88）%,P 〈 0.05] and induced cells apoptosis [（12.50 ± 0.66） % vs.（4.91 ± 0.15） %,P 〈 0.05].Conclusion The expression of miR-874 was low in glioblastoma.Loss of miR-874 is correlated with Tumorigenesis and development.The miR-874 might be a valid target for the diagnosis and treatment of glioblastoma.