Veterinary Archives, Vol. 91 No. 2, 2021.
Original scientific paper
https://doi.org/10.24099/vet.arhiv.1305
MiRNA-let-7b decreases proliferation activities and development of follicular cells via targeting MAP3K1 gene
Zhuandi Gong
; Hospital, Medicine College, Northwest Minzu University, Lanzhou, China
Yuhui Niu
; College of Life Science and Engineering, Northwest Minzu University, Lanzhou, China
Yuan Zhaofang
; College of Life Science and Engineering, Northwest Minzu University, Lanzhou, China
Juan Yang
; Biomedicine Research Center, Northwest Minzu University, Lanzhou, China
Suocheng Wei
; College of Life Science and Engineering, Northwest Minzu University, Lanzhou, China; Biomedicine Research Center, Northwest Minzu University, Lanzhou, China
Abstract
To date, it has not yet been determined if the apoptosis of follicular granulosa cells (FGCs) is mediated by miR- let-7b via MAP3K1. In the present study, FGCs were transfected with a miR-let-7b mimic at different doses (0, 40, 60, 80, 100 and 120 μM), and were allocated into the control group (CG), and MIM-1, MIM-2, MIM-3, MIM-4 and MIM- 5, respectively. Expression levels of miR-let-7b and mitogen-activated protein kinase kinase kinase 1 (MAP3K1) mRNAs and proteins were determined using RT-PCR and Western blots. Luciferase report assay was applied to verify the targeting relationship between miR-let-7b and MAP3K1. The results revealed that the proliferation activity of FGCs in the MIM-4 group was significantly lower than that of the CG and MIM-1 groups (P<0.05). The MiR-let-7b mimic obviously reduced expression levels of miR-let-7b of the FGCs. The largest reduction was found in MIM-4. Levels of MAP3K1 mRNAs and proteins in the MIM-3 and MIM-4 groups were lower than that of the CG (P<0.05 or P<0.01). Co-transfection of let-7b mimic significantly inhibited luciferase activity (P<0.05) as compared with the CG. In conclusion, miR-let-7b may obviously depress the cell viability and accelerate apoptosis of ovine FGCs. Higher doses of miR-let-7b mimic (80 μM and 100 μM) could significantly depress expressions of miR-let-7b mRNAs, MAP3K1 mRNAs and protein in ovine FGCs. MiR-let-7b promoted FGCs apoptosis by inhibiting the MAP3K1 gene.
Keywords
miRNA-let-7b; follicular granulosa cells; MAP3K1; protein; expression
Hrčak ID:
258462
URI
Publication date:
8.6.2021.
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