Effect of Mir-127-3p on Proliferation，Apoptosis，Migration and Invasion of Uveal Melanoma Cells via Targeting MAPK4
- VernacularTitle:MiR-127-3p靶向MAPK4对葡萄膜黑色素瘤细胞增殖、凋亡、迁移和侵袭的影响
- Author:
Li1 WEI
1
;
Hong-mei2 LIAN
2
;
Peng1 LIU
1
;
Xinghua1 LIU
1
;
Shu-yi3 WU
3
;
Meng-meng1 LIU
1
Author Information
1. Department of Pathology，Zhengzhou Third People′s Hospital，Zhengzhou 45000，China
2. Department of Anesthesia The third people′s Hospital Affiliated to Zhengzhou University ，Zhengzhou 45000，China
3. Central Laboratory，Zhengzhou Third People′s Hospital，Zhengzhou 450000，China
- Publication Type:Journal Article
- Keywords:
uveal melanoma, miR-127-3p, MAPK4, proliferation, migration, invasion
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2020;41(1):76-85
- CountryChina
- Language:Chinese
-
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.
