Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Dalian Medical University, Dalian, Liaoning Province 116044, China.
Department of Clinical Laboratory, the First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province 116011, China.
Reprod Biol. 2022 Dec;22(4):100706. doi: 10.1016/j.repbio.2022.100706. Epub 2022 Oct 31.
Polycystic ovary syndrome (PCOS) is an endocrine disorder that affects fertility in women of reproductive age, and a leading cause of anovulatory infertility. Ovarian granulosa cells are a major functional cell type in the ovary that undergo epithelial-to-mesenchymal transition (EMT) to initiate ovulation. Protein glycosylation, catalyzed by specific glycosyltransferases, has been implicated in reproductive events, such as embryo implantation, endometrial receptivity, and decidualization, etc. However, the relationship between glycosylation and EMT-mediated ovulation in PCOS is not well understood. To clarify the role of cobalt chloride (CoCl) and α-1,3/1,6-mannosyltransferase (ALG2) in PCOS, transwell assay, Real-time PCR, Western blot, immunofluorescence, and sphere formation assay were applied to assess cell migration, invasion, EMT, and stemness of ovarian granulosa cells. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the serum level of ALG2 in PCOS patients. We found that CoCl promoted the migration, invasion, EMT, and stemness of ovarian granulosa cells by downregulating the expression of ALG2. Upregulation of ALG2 rescued the effects of CoCl partially, and inhibited the EMT and stemness of ovarian granulosa cells by inactivating the Wnt/β-catenin signaling pathway. Moreover, the serum level of ALG2 was increased in patients with PCOS. Elevated ALG2, in combination with testosterone, represented better diagnostic value for PCOS as a multimarker than ALG2 or testosterone alone as a single marker. Thus, ALG2, downregulated by CoCl, was increased in PCOS patients which inhibited the EMT and stemness of ovarian granulosa cells by deactivating the Wnt/β-catenin signaling pathway. The combination of ALG2 and testosterone might thus act as a novel but promising biomarker for PCOS detection.
多囊卵巢综合征（PCOS）是一种影响育龄妇女生育能力的内分泌疾病，也是无排卵性不孕的主要原因。卵巢颗粒细胞是卵巢的主要功能细胞类型，它经历上皮间质转化（EMT）以启动排卵。由特定糖基转移酶催化的蛋白糖基化已被牵涉到生殖事件中，如胚胎着床、子宫内膜容受性和蜕膜化等。然而，糖基化与 PCOS 中 EMT 介导的排卵之间的关系尚不清楚。为了阐明氯化钴（CoCl）和α-1,3/1,6-甘露糖基转移酶（ALG2）在 PCOS 中的作用，我们应用 Transwell 测定、实时 PCR、Western blot、免疫荧光和球体形成测定来评估卵巢颗粒细胞的迁移、侵袭、EMT 和干性。酶联免疫吸附测定（ELISA）用于测量 PCOS 患者血清中 ALG2 的水平。我们发现，CoCl 通过下调 ALG2 的表达促进卵巢颗粒细胞的迁移、侵袭、EMT 和干性。ALG2 的上调部分挽救了 CoCl 的作用，并通过抑制 Wnt/β-catenin 信号通路抑制卵巢颗粒细胞的 EMT 和干性。此外，PCOS 患者的血清 ALG2 水平升高。与单独的 ALG2 或睾酮相比，升高的 ALG2 与睾酮联合作为多标记物对 PCOS 的诊断价值优于单独的 ALG2 或睾酮作为单一标记物。因此，下调的 ALG2 被 CoCl 上调，在 PCOS 患者中增加，通过去激活 Wnt/β-catenin 信号通路抑制卵巢颗粒细胞的 EMT 和干性。因此，ALG2 与睾酮的组合可能作为一种新的、有前途的 PCOS 检测生物标志物。
