Acta Veterinaria et Zootechnica Sinica ›› 2023, Vol. 54 ›› Issue (6): 2421-2435.doi: 10.11843/j.issn.0366-6964.2023.06.021

• ANIMAL BIOTECHNOLOGY AND REPRODUCTION • Previous Articles     Next Articles

Effect of miR-144-5p Targeting WNT5a on the Proliferation and Apoptosis of Goat Ovarian Granulosa Cells

LIU Jie1,2, XU Xiangping1,2, DENG Ming1,3,4, ZOU Xian1,2, JIANG Shengwei1,2, LIU Dewu1,3,4, LIU Guangbin1,3,4, SUN Baoli1,3,4, GUO Yongqing1,3,4, LI Yaokun1,3,4*   

  1. 1. College of Animal Science, South China Agricultural University, Guangzhou 510642, China;
    2. Guangdong Key Laboratory of Animal Breeding and Nutrition, State Key Laboratory of Livestock and Poultry Breeding, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
    3. National Engineering Research Center for Livestock and Poultry Breeding, South China Agricultural University, Guangzhou 510642, China;
    4. Guangdong Key Laboratory of Agricultural Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China
  • Received:2022-08-30 Online:2023-06-23 Published:2023-06-16

Abstract: The study aimed to investigate the effect of miR-144-5p targeting WNT5a on proliferation and apoptosis of goat granulosa cells. Ovarian granulosa cells from 2-year-old healthy Leizhou female goats were selected to overexpress and repress miR-144-5p and WNT5a, and the effects of miR-144-5p and WNT5a on granulosa cell proliferation and apoptosis were investigated by fluorescent quantitative PCR (real-time qPCR, RT-qPCR), Western blot, dual luciferase reporter gene, CCK-8 and other techniques. The results showed that the expression of miR-144-5p was significantly higher in large follicular granulosa cells than that in small follicular granulosa cells (P<0.01), and the expression of WNT5a was significantly lower in large follicular granulosa cells than that in small follicular granulosa cells (P<0.01); CCK-8 assay revealed that miR-144-5p inhibited the proliferation of GCs, and overexpression of miR-144-5p resulted in significant downregulation of mRNA and protein expression of the apoptosis suppressor gene BCL-2 (P<0.01) and significant upregulation of mRNA and protein expression of the pro-apoptotic gene BAX (P<0.01); Wnt/β-Catenin pathway gene CTNNB1 mRNA and β-Catenin protein levels were highly significantly reduced (P<0.01); In addition, miR-144-5p could target WNT5a to inhibit its expression. Meanwhile, CCK-8 assay results showed that WNT5a promoted the proliferation of GCs, and overexpression of WNT5a significantly upregulated the mRNA and protein levels of the BCL-2 (P<0.01), significantly downregulated the mRNA level (P<0.05) and protein level (P<0.01) of BAX, and the Wnt/β-Catenin pathway gene CTNNB1 mRNA and β-Catenin protein levels were significantly increased (P<0.01). In summary, in GCs, miR-144-5p targets and negatively regulates WNT5a expression, regulates the Wnt/β-Catenin pathway, and then upregulates the BAX/BCL-2 ratio to inhibit granulosa cell proliferation and promote apoptosis, which may have an effect on ovulation and lambing numbers in goats.

Key words: granulosa cells, miR-144-5p, WNT5a, goat, reproduction

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