METTL3 promotes proliferation of goat endometrial epithelial cells by regulating CTGF in an m6A-dependent manner

• Published in: Biology of reproduction 2023-06, Vol.108 (6), p.902-911
• Main Authors: Sun, Ya, Zhang, Xin-Cheng, Li, Meng-Die, Bu, Li-Ge, Wang, Bo, Li, Ting-Yue, Ding, Nai-Zheng, Ni, Hua
• Format: Article
• Language: English
• Subjects: 3' Untranslated regions; Animals; Cell proliferation; Cell Proliferation - genetics; Connective tissue growth factor; Connective Tissue Growth Factor - genetics; Connective tissues; CTGF; early pregnancy; Endometrium; Epigenetics; Epithelial cells; Epithelial Cells - metabolism; Estrogens; Female; goat uterus; Goats - genetics; m6A; Methyltransferase; Methyltransferases - genetics; Methyltransferases - metabolism; METTL3; mRNA; N6-methyladenosine; Pregnancy; Progesterone; RESEARCH ARTICLE; RNA, Messenger - metabolism; Uterus
• ISSN: 0006-3363; 1529-7268
• DOI: 10.1093/biolre/ioad029

N6-methyladenosine (m6A), an epigenetic modification on RNAs, plays an important role in many physiological and pathological processes. However, the involvement of m6A in goat uterus during early pregnancy remains largely unknown. In this study, we found that the total m6A level was increasing in goat uterus as early pregnancy progressed. Methyltransferase-like 3 (METTL3) is a core catalytic subunit of the m6A methyltransferase. We thus determined the expression and regulation of METTL3 in goat uterus. METTL3 was highly expressed in the luminal and glandular epithelia from day 16 (D16) to D25 of pregnancy, and it could be up-regulated by estrogen and progesterone in goat uterus and primary endometrial epithelial cells (EECs). In EECs, knockdown or overexpression of METTL3 resulted in a significant decrease or increase of cell proliferation, respectively. METTL3 knockdown reduced the m6A level of not only total RNA but also connective tissue growth factor (CTGF) mRNA. Luciferase assay suggested that METTL3 might target the potential m6A sites in the 3'untranslated region (3'UTR) of CTGF mRNA. Moreover, METTL3 positively regulated CTGF expression, and CTGF knockdown significantly counteracted the promoting effect of METTL3 overexpression on EEC proliferation. Collectively, METTL3 is dynamically expressed in goat uterus and can affect EEC proliferation by regulating CTGF in an m6A-dependent manner. Our results will lay a foundation for further studying the crucial mechanism of METTL3-mediated m6A modification in goat uterus during early pregnancy.