Investigation of the impacts of lncRNA PVT1 and microRNA-106a-5p in os-teosarcoma MG-63 cells and its mechanisms
Objective To investigate the effects of lncRNA PVT1 and miR-106a-5p on the ma-lignant behaviors of osteosarcoma cells and its molecular mechanisms.Methods Osteosarcoma MG-63 cells were divided into NC group,PVT1-siRNA group,miRNA inhibitor group,and co-transfection group.Cell proliferation ability was detected by CCK-8 assay,cell apoptosis was detected by flow cytometry,cellular reactive oxygen species(ROS)levels were measured by DCFH-DA method,gene expression was detected by real-time fluorescent quantitative polymer-ase chain reaction(RT-qPCR),and the expression levels of glutathione peroxidase 4(GPX4),ferritin heavy chain(FTH),heme oxygenase-1(HO1),and murine double minute 4(MDM4)were detected by Western blot.Results The results of CCK-8 assay showed that the prolifera-tive capacity of MG-63 cells was reduced at 12 h,24 h,48 h and 72 h after knockdown of ln-cRNA PVT1(P＜0.05),and the results of flow cytometry assay showed that apoptosis level as well as the ROS level of MG-63 cells were significantly increased after knockdown of lncRNA PVT1(P＜0.05),and that the results of the combination of the lncRNA PVT1 knockdown group were reversed by miR-106a-5p inhibitor relative to the lncRNA PVT1 knockdown group(P＜0.05).miR-106a-5p inhibitor reversed the results in the lncRNA PVT1 knockdown group(P＜0.05).RT-qPCR assay showed that miR-106a-5p gene expression increased and MDM4 gene expression was inhibited after knockdown of lncRNA PVT1(P＜0.05),and Western blot showed that lncRNA PVT1 inhibited MDM4 gene expression(P＜0.05),and lncRNA PVT1 inhibited MDM4 gene expression(P＜0.05).Western blot results showed that knockdown of lncRNA inhibited MDM4,HO1,GPX4 and FTH protein levels after PVT1(P＜0.05).Con-clusion In human osteosarcoma MG-63 cells,inhibition of lncRNA PVT1 expression can pro-mote the expression of miR-106a-5p,facilitate ferroptosis and apoptosis,and inhibit prolifera-tion.
万方数据
维普
