miR-182通过RAC1通路促进骨肉瘤HOS细胞凋亡

miR-182 promotes apoptosis of osteosarcoma HOS cells through RAC1 pathway

目的:检测miR-182在骨肉瘤HOS细胞中的表达,通过构建过表达miR-182的转染载体检测miR-182对骨肉瘤HOS细胞凋亡的影响。方法:RT-qPCR检测骨肉瘤HOS细胞miR-182表达。构建过表达miR-182的转染载体,进行转染后细胞共培养,RT-qPCR检测转染后miR-182表达,显微镜下观察细胞形态。Western blot分析miR-182对RAC1通路蛋白的作用,CCK-8法分析过表达miR-182对骨肉瘤HOS细胞生长的抑制情况,流式细胞术评估miR-182对骨肉瘤HOS细胞细胞周期及凋亡的影响。结果:骨肉瘤HOS细胞中miR-182呈低表达。构建过表达miR-182的转染载体,骨肉瘤HOS细胞中RAC1通路蛋白及下游PAK1蛋白表达显著下调。过表达miR-182显著促进骨肉瘤细胞凋亡并阻滞其细胞周期进程。结论:miR-182可通过靶向调控RAC1通路促进骨肉瘤HOS细胞凋亡并阻止其细胞周期进程。

Objective:To detect expression level of miR-182 in osteosarcoma HOS cells,and to detect effect of miR-182 on apoptosis of osteosarcoma HOS cell by constructing an overexpression miR-182 transfection vector.Methods:miR-182 expression in osteosarcoma HOS cells was detected by RT-qPCR.A transfection vector of over-expressed miR-182 was constructed and cells were cocultured after transfection.miR-182 expression after transfection was detected by RT-qPCR,and cell morphology was observed under microscope.Western blot was performed to explore effect of miR-182 on RAC1 pathway proteins.CCK-8 was used to analyze effect of miR-182 over-expression on proliferation activity of osteosarcoma HOS cells.Effects of miR-182 over-expression on cell cycle and apoptosis of osteosarcoma HOS cells were evaluated by flow cytometry.Results:miR-182 was lower expressed in osteosarcoma HOS cells.After over-expressed miR-182 transfection vector was constructed,expressions of RAC1 pathway proteins and downstream PAK1 proteins in osteosarcoma HOS cells were significantly decreased.miR-182 over-expression significantly promoted apoptosis of osteosarcoma cells and blocked their cell cycle progression.Conclusion:miR-182 can promote apoptosis and block cell cycle progression of osteosarcoma HOS cells through targeted regulation of RAC1 pathway.