Fu Bingxin, Guo Manman, Jia Yuanyuan, Han Xiaoyu, Bi Beibei, Fang Lanlan, Cheng Jung-Chien
Center for Reproductive Medicine, Henan Key Laboratory of Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China.
Center for Reproductive Medicine, Henan Key Laboratory of Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China.
Mol Cell Endocrinol. 2025 Sep 15;607:112604. doi: 10.1016/j.mce.2025.112604. Epub 2025 Jun 10.
Amphiregulin (AREG), an epidermal growth factor (EGF)-like ligand, is the predominant epidermal growth factor receptor (EGFR) ligand in human follicular fluid and mediates the effects of luteinizing hormone (LH) on ovarian function. In this study, we investigated whether AREG regulates the expression of cysteine-rich angiogenic inducer 61 (CYR61) and connective tissue growth factor (CTGF), two key matricellular proteins involved in ovarian function, and whether they mediate AREG-induced cyclooxygenase-2 (COX-2) expression. Using the human granulosa cell tumor-derived KGN cell line and primary human granulosa-lutein (hGL) cells, we demonstrated that AREG treatment upregulated CYR61 and CTGF protein levels in an EGFR-dependent manner. Mechanistic analysis revealed that AREG-induced expression of CYR61 and CTGF was mediated through the ERK1/2, AKT, CREB, and YAP signaling pathways. Inhibition of these pathways using specific inhibitors or small interfering RNA blocked AREG-induced CYR61 and CTGF expression, indicating their essential roles in this process. Moreover, knockdown of CYR61 and CTGF attenuated AREG-induced COX-2 expression, establishing their role as key mediators of AREG signaling in human granulosa cells. Finally, our results showed that LH treatment induced the expression of CYR61 and CTGF, and this induction was attenuated by EGFR inhibition. Moreover, knockdown of CYR61 and CTGF reduced LH-induced COX-2 expression. These findings provide novel insights into the molecular mechanisms by which AREG regulates ovarian function and highlight potential targets for reproductive health research.
双调蛋白（AREG）是一种表皮生长因子（EGF）样配体，是人类卵泡液中主要的表皮生长因子受体（EGFR）配体，并介导促黄体生成素（LH）对卵巢功能的影响。在本研究中，我们调查了AREG是否调节富含半胱氨酸的血管生成诱导因子61（CYR61）和结缔组织生长因子（CTGF）的表达，这两种关键的基质细胞蛋白参与卵巢功能，以及它们是否介导AREG诱导的环氧合酶-2（COX-2）表达。使用人颗粒细胞瘤衍生的KGN细胞系和原代人颗粒黄体（hGL）细胞，我们证明AREG处理以EGFR依赖性方式上调CYR61和CTGF蛋白水平。机制分析表明，AREG诱导的CYR61和CTGF表达是通过ERK1/2、AKT、CREB和YAP信号通路介导的。使用特异性抑制剂或小干扰RNA抑制这些通路可阻断AREG诱导的CYR61和CTGF表达，表明它们在这一过程中的重要作用。此外，敲低CYR61和CTGF可减弱AREG诱导的COX-2表达，确立了它们作为人类颗粒细胞中AREG信号关键介质的作用。最后，我们的结果表明，LH处理可诱导CYR61和CTGF的表达，而EGFR抑制可减弱这种诱导作用。此外，敲低CYR61和CTGF可降低LH诱导的COX-2表达。这些发现为AREG调节卵巢功能的分子机制提供了新的见解，并突出了生殖健康研究的潜在靶点。
