Characterization of miRNA-483-5p and targeted gene ERK1 in the regulation of proliferation-apoptosis balance of human granulosa cells
- VernacularTitle:miR-483-5p 通过靶基因 ERK1调控人类颗粒细胞增殖凋亡平衡
- Author:
Xiumin JIANG
;
Yusheng LIU
;
Bo XU
- Publication Type:Journal Article
- Keywords:
miR-483-5p;
ERK1 /2-MAPK pathway;
granular cells;
proliferation-apoptosis balance
- From:
Acta Universitatis Medicinalis Anhui
2015;(11):1639-1644
- CountryChina
- Language:Chinese
-
Abstract:
Objective To demonstrate that miR-483-5p and its targeted gene ERK1 involved in the regulation of proliferation and apoptosis of human granulosa cells.Methods ① The ERK1 mRNA and protein level expression were detected by PCR and Western blot after miR-483-5p overexpression in vitro normal human granular cells.De-tected the relative luciferase density after cotransfecting miR-483-5p mimics and its control respectively with wild or mutant ERK1 mRNA 3`UTR cloned luciferase report vector in granular cells.② Granular cells proliferation and ap-optosis were detected by MTT and TUNEL after transfecting miR-483-5p mimics or ERK1 siRNAs alone or simulta-neously.Results ① It showed that both ERK1 mRNA and protein in granular cells were markedly downregulated after the transfection of miR-483-5p mimics.A significant relative luciferase activity decrease were detected in the granular cells co-transfected with ERK1 wild and miR-483-5p mimics comparison with the control mimics,but not in the cells co-transfected with the ERK1 mutant and miR-483-5p mimics.② When miR-483-5p mimics and ERK1 siRNAs were alone or co-transfected into granular cells,the proliferation was inhibited while apoptosis trend was monitored to be promoted in all cases.No obvious statistic difference was shown between each other.Conclusion MiR-483-5p is involved in the regulation of the proliferation and apoptosis of human granulosa cells by directly binding to the target gene ERK1.
