Cao Hui, Li Ranran, Ling Li, Ni Guantai
Anhui Medical University, Hefei, Anhui, 230000, PR, China.
Department of Obstetrics and Gynaecology, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui, 241000, PR, China.
Endocr Metab Immune Disord Drug Targets. 2024 Dec 17. doi: 10.2174/0118715303299470240723060939.
Recurrent Miscarriage (RM) is a chronic and heterogeneous autoimmune disease. Platelet factor 4 (PF4) has been found to be involved in the pathogenesis of RM.
We aimed to explore the role and mechanism of PF4 on trophoblasts in RM in vitro.
In this study, the expression of PF4 and PF4 receptor CXC chemokine receptor 3 (CXCR3) in the placentas of patients with RM were analyzed by RT-qPCR and western blotting. Serum PF4 level was tested by ELISA. Following PF4 silencing and SOCS3 overexpression in HTR-8/SVneo cells, cell proliferation was detected by CCK-8, colony formation, and EDU assays. Wound healing and transwell assays separately evaluated cell migration and invasion. Immunofluorescence assay determined E-cadherin expression. Tube formation assay was used to measure the angiogenesis. Western blotting examined the expression of metastasis, epithelial-mesenchymal transition (EMT) and suppressor of cytokine signaling 3 (SOCS3)/signal transducer and activator of transcription 3 (STAT3) signaling-associated proteins.
The results revealed that PF4 displayed increased expression in placental villus tissues of RM patients. Serum PF4 level was also elevated in RM patients. PF4 silencing promoted the proliferation, migration, invasion, EMT, and angiogenesis of HTR-8/SVneo cells. Additionally, PF4 knockdown downregulated SOCS3 expression to activate STAT3 signaling. SOCS3 overexpression countervailed the effects of PF4 deficiency on HTR-8/SVneo cells.
In summary, PF4 participated in the proliferation, migration, invasion, EMT and angiogenesis of trophoblast cells by modulating the SOCS3/STAT3 signaling pathway, indicating that targeting the PF4/SOCS3/STAT3 pathway could be a novel therapy for RM.
复发性流产（RM）是一种慢性异质性自身免疫性疾病。已发现血小板因子4（PF4）参与RM的发病机制。
我们旨在探讨PF4在体外对RM患者滋养层细胞的作用及机制。
在本研究中，通过RT-qPCR和蛋白质印迹法分析RM患者胎盘组织中PF4及其受体CXC趋化因子受体3（CXCR3）的表达。采用酶联免疫吸附测定法检测血清PF4水平。在HTR-8/SVneo细胞中沉默PF4并过表达细胞因子信号转导抑制因子3（SOCS3）后，通过CCK-8法、集落形成实验和5-乙炔基-2'-脱氧尿苷（EDU）实验检测细胞增殖情况。划痕实验和Transwell实验分别评估细胞迁移和侵袭能力。免疫荧光实验检测E-钙黏蛋白的表达。采用管腔形成实验检测血管生成情况。蛋白质印迹法检测转移、上皮-间质转化（EMT）以及SOCS3/信号转导和转录激活因子3（STAT3）信号相关蛋白的表达。
结果显示，PF4在RM患者胎盘绒毛组织中的表达增加。RM患者血清PF4水平也升高。沉默PF4可促进HTR-8/SVneo细胞的增殖、迁移、侵袭、EMT和血管生成。此外，敲低PF4可下调SOCS3表达以激活STAT3信号通路。过表达SOCS3可抵消PF4缺乏对HTR-8/SVneo细胞的影响。
综上所述，PF4通过调节SOCS3/STAT3信号通路参与滋养层细胞的增殖、迁移、侵袭、EMT和血管生成，表明靶向PF4/SOCS3/STAT3通路可能是RM的一种新疗法。
