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Article in Chinese | WPRIM | ID: wpr-817635

ABSTRACT

@#【Objective】To investigate the effect of miR- 127-3p on proliferation,apoptosis,migration and invasion of uveal melanoma cells.【Methods】The expression of miR- 127- 3p and MAPK4 mRNA in human uveal melanoma tissues and cells,normal tissues and cells were detected by RT-qPCR. The mimic-NC,miR-127-3p mimic,pc-MAPK4 plasmids were transfected into SP6.5 or OM431 cells,respectively,by Lipofectamine 2000. The relationship between miR-127-3p and MAPK4 counterstaining was detected by dual luciferase assay. Cell proliferation was detected by CCK- 8 method,apoptosis was detected by flow cytometry,cell migration ability was detected by scratch test,cell invasion ability was detected by Transwell method,and relative expression level of AKT/mTOR pathway protein was detected by Western blot.【Results】In uveal melanoma tissues and cell lines,the expression of miR- 127-3p was down-regulated(P < 0.01)while that of MAPK4 expression was significantly up-regulated(P < 0.01). The binding site of miR-127-3p and MAPK4 3′UTR region,the high expression of miR-127-3p significantly inhibited the luciferase activity of wild-type MAPK4 plasmid(P < 0.01),but the mutant MAPK4 plasmid Luciferase activity has no effect. Compared with the Control group ,the proliferation of SP6.5 cells and OM431 cells in miR- 127-3p mimic group were significantly decreased(P < 0.01),and the apoptotic rate was significantly increased(P < 0.01). The scratch closure rate was obvious. The decrease(P < 0.01), the number of invading cells per field was significantly decreased(P < 0.01),and the expression of p-AKT(T308)/AKT and p-mTORr(S473)/mTOR protein were significantly down-regulated(P < 0.01). Transfection of pc-MAPK4 reversed the above changes.【Conclusion】MiR-127-3p inhibits proliferation,migration and invasion of uveal melanoma cells and induces apoptosis by down-regulating MAPK4,which may be involved in the inhibition of AKT/mTOR pathway activation.

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