miR-192在胶质瘤U251细胞中的表达及对其增殖、迁移及凋亡能力的影响

Expression of miR-192 in Glioma U251 Cells and Its Effect on Proliferation,Migration and Apoptosis

目的探讨miR-192在胶质瘤U251细胞中的表达及对其增殖、迁移及凋亡等生物学能力的影响。方法采用RT-PCR检测胶质瘤细胞株U251、LN18、U373和正常人脑胶质细胞株中HEB的表达水平。采用脂质体转染法将miR-192类似物(miR-192 mimics)和阴性对照(miR-negtive control,miR-NC)分别转染U251细胞。应用CCK-8法和平板克隆形成实验检测细胞增殖能力的变化情况。应用Transwell实验检测细胞迁移能力的变化,应用流式细胞术检测细胞凋亡情况的变化。应用生物信息学方法预测miR-192的靶基因。结果miR-192在胶质瘤细胞株U251、LN18和U373中相对表达水平均低于HEB组,差异有统计学意义(P<0.05)。miR-192 mimics和miR-NC分别转染U251细胞,CCK-8实验显示:miR-192 mimics组在第4、5天时的OD值明显低于miR-NC组,差异有统计学意义(P<0.05)。平板克隆形成实验显示:miR-192 mimics组细胞克隆形成率为(28.23±0.81)%,明显低于miR-NC组[(41.59±1.21)%],差异有统计学意义(P<0.05)。Transwell实验结果显示,转染miR-192 mimics的穿膜细胞数为(83.20±21.51)个/孔,明显少于miR-NC组[(126.33±24.50)个/孔],差异有统计学意义(P<0.05)。流式细胞术检测显示,miR-192 mimics组和miR-NC组的细胞凋亡率分别为(18.23±1.86)%和(10.43±1.40)%,miR-192 mimics组的细胞凋亡率明显高于miR-NC组,差异有统计学意义(P<0.05)。生物信息学方法预测SMC5可能是miR-192的靶基因。结论miR-192能够抑制胶质瘤U251细胞的增殖、迁移并促进其凋亡,是胶质瘤的治疗提供潜在靶点。miR-192可能通过靶基因SMC5发挥其对胶质瘤的调控作用。

Objective To investigate the expression of miR-192 in glioma U251 cells and its effect on biological ability such as proliferation,migration and apoptosis.Methods RT-PCR was used to detect the expression levels of glioma cell lines U251,LN18,U373 and normal human brain glioma cell lines HEB.The liposome transfection method was used to transfect miR-192 analogs(miR-192 mimics)and negative control(miR-negtive control,miR-NC)to U251 cells,respectively.The CCK-8 method and plate cloning assay were used to detect changes in cell proliferation.Transwell assay was used to detect changes in cell migration ability,and flow cytometry was used to detect changes in apoptosis.Using bioinformatics to predict the target genes of miR-192.Results The relative expression levels of miR-192 in glioma cell lines U251,LN18 and U373 were lower than those in the HEB group,and the difference was statistically significant(P<0.05).miR-192 mimics and miR-NC were transfected into U251 cells,respectively.The CCK-8 experiment showed that the OD value of the miR-192 mimics group on the 4th and 5th days was significantly lower than that of the miR-NC group,and the difference was statistically significant(P<0.05).Plate colony formation experiments showed that the colony formation rate of miR-192 mimics group was(28.23±0.81)%,which was significantly less than that of miR-NC group[(41.59±1.21)%],and the difference was statistically significant(P<0.05).Transwell experiments showed that the number of transmembrane cells transfected with miR-192 mimics was(83.20±21.51)cells/well,which was significantly less than the miR-NC group[(126.33±24.50)cells/well].The difference was statistically significant(P<0.05).Flow cytometry showed that the apoptotic rates of miR-192 mimics group and miR-NC group were(18.23±1.86)%and(10.43±1.40)%,respectively.The apoptosis rate of miR-192 mimics group was significantly higher than that of miR-NC group,and the difference was statistically significant(P<0.05).Bioinformatics methods predict that SMC5 may be a target gene of miR-192.Conclusion miR-192 can inhibit the proliferation,migration and promote apoptosis of glioma U251 cells,and is a potential target for the treatment of gliomas.miR-192 may exert its regulatory effect on glioma through the target gene SMC5.