Effect of miR-519e-5p targeting ANXA5 on the biological behavior of chordoma U-CH1 cells
Objective To investigate the effect of miR-519e-5p targeting Annexin A5(ANXA5)on the biological behavior of chordoma U-CH1 cells.Methods The expressions of miR-519e-5p and ANXA5 in human chordoma tissues and adjacent tissues were detected by qRT-PCR.U-CH1 cells were cultured in vitro and divided into miR-519e-5p mimics group,miR-519e-5p mimics negative control+empty vector group,miR-519e-5p mimics+ANXA5 overexpression group,and control group.The proliferation of U-CH1 cells in each group was detected by plate colony formation assay and MTT assay;flow cytometry was used to detect the apoptosis of U-CH1 cells in each group;the migration and invasion of U-CH1 cells in each group were detected by scratch would healing assay and Transwell assay respectively;the expression of ANXA5 protein,apoptosis-related proteins(Bcl-2,Bax)and migration epithelial-mesenchymal transition-related proteins(Claudin-1,Vimentin,E-cadherin)of U-CH1 cells in each group were detected by Western blot;the expression of miR-519e-5p and ANXA5 mRNA of U-CH1 cells in each group was detected by qRT-PCR;and the interaction between miR-519e-5p and ANXA5 in U-CH1 cells was examined by dual-luciferase reporter gene assay.Results Compared with the adjacent tissues,the expression of miR-519e-5p in chordoma tissues decreased(P<0.05),and the expression of ANXA5 mRNA increased(P<0.05).Compared with the control group,the cell proliferation rate,colony formation rate,migration rate,invasion number,expressions of ANXA5 mRNA and ANXA5,Bcl-2,Claudin-1,and Vimentin proteins in the miR-519e-5p mimics group decreased(P<0.05),the expression of miR-519e-5p mRNA,the apoptosis rate,and the expression of Bax and E-cadherin proteins increased(P<0.05).Compared with the miR-519e-5p mimics group,the colony formation rate,cell proliferation rate,invasion number,migration rate,expressions of ANXA5 mRNA and ANXA5,Bcl-2,Vimentin,and Claudin-1 proteins in the miR-519e-5p mimics+ANXA5 overexpression group increased(P<0.05),and the apoptosis rate,the expression of Bax and E-cadherin proteins decreased(P<0.05).MiR-519e-5p can target and down-regulate the expression of ANXA5 in U-CH1 cells.Conclusion MiR-519e-5p reduces the migration,proliferation and invasion viability of chordoma U-CH1 cells,and promotes apoptosis by targeting the inhibition of ANXA5 expression.
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维普
