目的:反复植入失败(RIF)患者子宫内膜增殖细胞核抗原(PCNA)的表达是否调节子宫内膜容受性?
结论:高丰度的PCNA减弱RIF患者的子宫内膜粘附能力和蜕膜化。
背景:已在多种不育相关疾病中发现了PCNA的异常表达。然而,PCNA在RIF患者子宫内膜容受性和子宫内膜蜕膜化中的表达模式和作用尚不清楚。
方法:我们分析了24例RIF患者和24例健康女性子宫内膜分泌中期组织中PCNA的表达。此外,在月经周期正常的妇女的增殖期和分泌中期子宫内膜组织样本中以及在正常早期妊娠期间从10名妇女的蜕膜组织样本中测量PCNA表达水平(每组n=10)。使用BeWo球体研究PCNA在子宫内膜粘附能力和子宫内膜蜕膜化中的功能和调节机制,石川细胞,和人子宫内膜基质细胞(HESC)。
方法:用RT-qPCR方法分析RIF患者和正常子宫内膜妇女子宫内膜中期分泌组织、月经周期不同阶段子宫内膜组织和蜕膜化组织中PCNA的表达,westernblot,免疫组织化学染色(IHC)。此外,直接附着在Ishikawa细胞单层上的BeWo球体的数量,以及潜在的分子机制,在过表达PCNA的细胞和对照组之间进行比较。此外,研究了PCNA对HESCs蜕膜化的影响及其调控机制。
结果:我们的研究结果表明,RIF患者子宫内膜中部分泌组织中PCNA的丰度明显高于子宫内膜健康的女性。PCNA的表达在月经周期的增殖阶段增加,但在分泌中期和蜕膜组织中逐渐降低。有趣的是,PCNA在人子宫内膜上皮细胞(HEEC)和HESC中均有表达。在石川牢房里,PCNA过表达通过抑制粘附分子(E-钙黏着蛋白和整联蛋白β3)的表达和激活FAK/桩蛋白信号通路而显著降低子宫内膜粘附能力。此外,在HESCs中,PCNA过表达通过激活AKT/β-连环蛋白信号通路并通过上调ZO-1和occludin表达来增加细胞之间的紧密连接来减弱子宫内膜蜕膜化。此外,PCNA-ELAVL1相互作用通过在蜕膜化HESC中的共免疫沉淀来确认。
方法:不适用。
结论:PCNA的功能分析受到人子宫内膜组织数量的限制。需要更大的样本量来进一步探索PCNA在胚胎植入过程中的潜在作用。此外,目前的结果应该谨慎对待,因为只有少数在RIF患者人群中移植的胚胎接受了胚胎染色体非整倍体(PGT-A)的植入前遗传学检测,尽管胚胎倍性测试对无法解释的RIF的诊断具有重要意义。
结论:高PCNA表达减弱RIF患者的子宫内膜粘附能力和蜕膜化。这些发现为植入失败发生的潜在机制提供了新的见解。
背景:这项工作得到了国家自然科学基金(82101698)的支持,山东省自然科学基金(ZR2021MH012),《烟台市科技计划》(2023YD021、2022YD031)。作者没有利益冲突要披露。
背景:不适用。
OBJECTIVE: Does the expression of proliferating cell nuclear antigen (PCNA) in the endometrium regulate endometrial receptivity in patients with recurrent implantation failure (RIF)?
CONCLUSIONS: A high abundance of PCNA attenuates endometrial adhesive capacity and decidualization in patients with RIF.
BACKGROUND: Aberrant expression of PCNA has been discovered in multiple infertility-related disorders. However, the expression pattern and role of PCNA in the establishment of endometrial receptivity and endometrial decidualization in patients with RIF remain unclear.
METHODS: We analysed the expression of PCNA in mid-secretory endometrial tissues from 24 patients with RIF and 24 healthy women. Additionally, PCNA expression levels were measured in proliferative and mid-secretory phase endometrial tissue samples from women with regular menstrual cycles and in decidual tissue samples taken from ten women during normal early pregnancy (n = 10 per phase for each group). The function and regulatory mechanisms of PCNA in endometrial adhesive capacity and endometrial decidualization were investigated using BeWo spheroids, Ishikawa cells, and human endometrial stromal cells (HESCs).
METHODS: The expression of PCNA in mid-secretory endometrial tissues of patients with RIF and women with normal endometrium and in endometrial tissue at different stages of the menstrual cycle and in decidualized tissues was analysed by RT-qPCR, western blot, and immunohistochemistry staining (IHC). Furthermore, the number of BeWo spheroids directly attached to the Ishikawa cell monolayers, and the potential molecular mechanisms involved, were compared between cells overexpressing PCNA and a control group. Additionally, the effect and regulatory mechanisms of PCNA on the decidualization of HESCs in vitro were investigated.
RESULTS: Our findings indicated that the abundance of PCNA was dramatically greater in mid-secretory endometrial tissues from patients with RIF than in those from women with healthy endometrium. The expression of PCNA increased in the proliferative phase of the menstrual cycle but decreased gradually in the mid-secretory phase and in decidual tissues. Interestingly, PCNA was expressed in both human endometrial epithelial cells (HEECs) and HESCs. In Ishikawa cells, PCNA overexpression dramatically reduced the endometrial adhesive capacity by inhibiting the expression of adhesion molecules (E-cadherin and integrin β3) and activating the FAK/paxillin signalling pathway. Furthermore, in HESCs, PCNA overexpression attenuated endometrial decidualization by activating the AKT/β-catenin signalling pathway and increasing tight junctions between cells by upregulating ZO-1 and occludin expression. In addition, PCNA-ELAVL1 interactions were confirmed by coimmunoprecipitation in decidualized HESCs.
METHODS: N/A.
CONCLUSIONS: The functional analysis of PCNA was limited by the number of human endometrial tissues. A larger sample size is required to further explore the potential roles of PCNA during embryo implantation. Moreover, the present results should be taken with caution, as only a few of the embryos that were transferred in RIF patients population underwent preimplantation genetic testing for embryonic chromosome aneuploidies (PGT-A), despite embryo ploidy testing being significant in the diagnosis of unexplained RIF.
CONCLUSIONS: High PCNA expression attenuates endometrial adhesive capacity and decidualization in patients with RIF. These findings provide new insights into the potential mechanisms underlying the occurrence of implantation failure.
BACKGROUND: This work was supported by the National Natural Science Foundation of China (82101698), Shandong Provincial Natural Science Foundation (ZR2021MH012), and the Science and Technology Plan of Yantai (2023YD021 and 2022YD031). The authors have no conflicts of interest to disclose.
BACKGROUND: N/A.