miR-155抑制物对骨肉瘤细胞增殖与凋亡的影响

摘要

Objective:To investigate the effect of microRNA-155 inhibitors on the biological behaviour of osteosarcoma cells by observing the role of microRNA-155 inhibitors in cell proliferation and apoptosis of human osteosarcoma cell line SOSP-9607.Methods:The experiment contained three groups:experimental group (transfected with hsa-miR-155 inhibitors),control group (transfected with microRNA inhibitor N.C) and blank group.Detected the MTF value (OD) at 24h,48h,72h and 96h,then showed the cell growth curve.The apoptosis and cell cycle were measured by flow cytometry.Results:The experimental group showed lower cell proliferation of efficiency than the control group,and there's no obvious effect between the control group and blank group.There's no difference between control group and blank group,while the apoptosis ratio of the experimental group [(8.5 ± 0.4) ％] was much higher than that of the control group [(1.2 ± 0.3) ％] and blank group [(1.1 ± 0.2) ％] (P ＜ 0.01).In comparison with the control group and blank group,the experimental group increased the percentage of cells at G0/G1 phase and decreased the percentage of cells G2/M phase and S phase (P ＜ 0.01).Conclution:The hsa-miR-155 inhibitors may have a great effect on the proliferation,apoptosis and cell cycle arrest of human osteosarcoma SOSP-9607 cells by down-regulating miR-155.%目的:观察miR-155抑制物对于骨肉瘤细胞系SOSP-9607细胞增殖和凋亡的影响,探讨miR-155抑制物在骨肉瘤细胞生物学行为中所起的作用.方法:实验分3组,分别为实验组、阴性对照组和正常细胞对照组.实验组转染miR-155抑制物(hsa-miR-155 inhibitors)抑制其细胞内miR-155活性,阴性对照组转染抑制物阴性对照核苷酸序列(microRNA inhibitor N.C),正常细胞对照组不做转染.采用四甲基偶氮唑蓝(MTT)比色实验测定3组细胞24小时、48小时、72小时和96小时的吸光度(OD)值,并绘制生长曲线.细胞周期与凋亡的测定采用流式细胞仪检测.结果:两个对照组细胞增殖速度无明显差异,而实验组细胞增殖速度则明显减慢.两对照组细胞凋亡率未见明显差异,而实验组细胞凋亡率(8.5±0.4)％与阴性对照组凋亡率(1.2±0.3)％、空白对照组凋亡率(1.1±0.2)％相比则明显增高(P＜O.O1).相比于对照组,实验组G0/G1期细胞所占比例明显增加,与此同时G2/M期与S期细胞比例则显著减少(P＜0.01).结论:miR-155抑制物可通过抑制成骨肉瘤细胞系SOSP-9607细胞内miR-155的活性,显著抑制其增殖能力,促进细胞凋亡和细胞G0/G1期阻滞.miR-155可能会成为骨肉瘤诊治的新靶点.