Molecular mechanism of miR-101-3p reversing docetaxel resistance in breast cancer through targeted downregulation of GALNT1
Objective To explore the molecular mechanism of miR-101-3p reversing docetaxel(DTX)resistance in breast cancer through targeted downregulation of GALNT1.Methods MCF-7/DTX cell line was constructed,and the expression level of miR-101-3p in MCF-7,MCF-7/DTX cell lines and cancer tissue were detected by qRT-PCR.MCF-7/DTX cells were transfected with miR-101-3p mimics,siGALNT1 and miR-101-3p inhibitor,then the transfection efficiency was assessed by qRT-PCR assay.The effects of regulating miR-101-3p and GALNT1 on DTX sensitivity of MCF-7/DTX cells was detected by MTT.The changes in the migration ability of each transfected groups were analyzed by wound healing assay.Changes in vertical invasion ability of MCF-7/DTX cells after regulating miR-101-3p and GALNT1 were discovered by transwell assay.The percentage of apoptosis of cells in each group under the same concentration of DTX treatment was detected by flow apoptosis assay.The effects of regulating miR-101-3p and GALNT1 on the expression of apoptosis-related proteins and EMT marker proteins was evaluated by western blot.Results The relative expression of miR-101-3p in MCF-7/DTX cells and DTX resistant group showed an abnormal decrease trend.After transfection with miR-101-3p mimics,the DTX sensitivity of the miR-101-3p mimics group was significantly increased compared with the miR-NC group.The ability of migration and invasion decreased at high levels of miR-101-3p,while the percentage of apoptosis increased significantly.Apoptotic protein expression showed the same trend,and the process of epithelial-mesenchymal transition was inhibited to a certain extent.The relative expression of GALNT1 in MCF-7/DTX cells was significantly higher than that in MCF-7 cells.Dual-luciferase experiments confirmed the targeting relationship between miR-101-3p and GALNT1,and the expression of GALNT1 was negatively correlated with miR-101-3p.The rescue experiment further analyzed the relationship between GALNT1 and miR-101-3p.Low levels of miR-101-3p could reverse the DTX hypersensitivity caused by knockdown of GALNT1,and at the same time,the changes of migration,invasion,apoptosis and EMT of MCF-7/DTX cells induced by knockdown of GALNT1 were restored to a certain extent.Conclusion miR-101-3p-mediated DTX sensitivity of MCF-7/DTX cells may be accomplished by targeting GALNT1.
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