miRNA-21抑制剂对骨肉瘤MG63细胞增殖、凋亡和迁移的影响

摘要

目的：通过观察miR-21抑制剂对骨肉瘤MG63细胞增殖、凋亡及迁移能力带来的变化，探讨miR-21抑制剂在骨肉瘤细胞生物学行为中的作用。方法：将MG63细胞随机分为实验组、阴性对照组和正常细胞对照组。实验组转染miR-21抑制剂（hsa-miR-21 inhibitors）抑制其细胞内miR-21活性，阴性对照组转染抑制剂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MTT比色实验测定三组细胞24h、48h、72h和96h的吸光度（OD）值，并绘制生长曲线。采用流式细胞仪检测细胞凋亡。Tran-swell法检测细胞迁移。结果：两对照组间细胞的增殖速度无明显差异（P＞0．05），与对照组相比，实验组细胞增殖速度明显减慢（P＜0．05）。实验组细胞凋亡率明显增高（P＜0．05）；阴性对照组、空白对照组之间细胞凋亡无明显差异（P＞0．05）。实验组细胞迁移力受到明显抑制（P＜0．05）。与空白对照组相比，阴性对照组细胞迁移力无明显差异（P＞0．05）。结论：miR-21抑制剂可通过抑制骨肉瘤细胞系MG63细胞内miR-21的活性，显著抑制其增殖能力，促进细胞凋亡和降低迁移。miR-21可能成为骨肉瘤诊治的新靶点。%Objective:To investigate the effect of microRNA-21 inhibitors on the biological behavior of osteosar-coma cells by observing the role of microRNA-21 inhibitors in cell proliferation,apoptosis and migration of human osteosarcoma cell line MG63.Methods:MG63 cells were randomly divided into three groups:Experimental group (transfected with hsa-miR-21 inhibitors),control (transfected with microRNA inhibitor N.C)group and blank group.Detected the MTT value (OD)at 24h,48h,72h and 96h,then showed the cell growth curve.The apoptosis was measured by flow cytometry.Cell number of migration was measured by Transwell assay.Results:There's no obvious effect between the control group and blank group (P>0.05),and the experimental group showed lower cell prolifera-tion of efficiency than the control group (P<0.05).There's no difference between control group and blank group (P>0.05),while the apoptosis ratio of the experimental group was much higher than that of the control group and blank group (P<0.05).The migration of the experimental group cells was significantly inhibited (P<0.05).Conclusion:hsa-miR-21 inhibitors may have a great effect on the proliferation,apoptosis and migration arrest of human osteo-sarcoma MG63 cells by down-regulating miR-21.miR-21 may become a new target in the diagnosis and treatment of osteosarcoma.