Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200135, China; Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China.
Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200135, China; Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China.
Placenta. 2020 Jan 1;89:33-41. doi: 10.1016/j.placenta.2019.10.007. Epub 2019 Oct 18.
Embryo implantation depends on trophoblast cells migration and invasion. Abnormal function of trophoblast cells could result in many pregnancy complications. Secreted protein acidic and rich in cysteine like-1 (SPARCL1) has been reported to inhibit cell migration and tumor invasion. This study aimed to explore the role of SPARCL1 in trophoblast functions.
Villous specimens were obtained from 31 women with spontaneous abortion and 31 women with normal early pregnancy to determine the expression of SPARCL1. HTR8/SVneo cells and JAR cells were transfected with pIRES2-EGFP-SPARCL1 vectors and control vectors. The proliferation assay and scratch-wound assay were performed. Quantitative polymerase chain reaction (qPCR) and western blotting were performed to assess epithelial mesenchymal transition (EMT)-related molecules including MMP2, MMP3, N-cadherin, E-cadherin and vimentin. Extracellular signal-regulated kinase (ERK) phosphorylation activity and AP-1 expression in HTR8/SVneo cells following multi-scratching were detected using above assays.
The mRNA and protein levels of SPARCL1 were significantly higher in the abortion group than in the normal pregnancy group. After transfection, there was no difference of cell viability between the SPARCL1-overexpression group and control vector group. However, the migration distance and area were reduced and the abundances of EMT related molecules were changed by SPARCL1 overexpression when compared with controls. Lower ERK phosphorylation activity and decreased Fos and Jun expressions were noted at high level of SPARCL1.
Restrained migration and invasion were noted in trophoblast cells with SPARCL1 overexpression, which might affect embryo implantation and placenta development. It could be involved in the pathogenesis of spontaneous abortion.
胚胎着床依赖于滋养层细胞的迁移和侵袭。滋养层细胞功能异常可导致多种妊娠并发症。富含半胱氨酸的酸性分泌蛋白 1（SPARCL1）已被报道可抑制细胞迁移和肿瘤侵袭。本研究旨在探讨 SPARCL1 在滋养层功能中的作用。
从 31 例自然流产和 31 例正常早孕的妇女中获取绒毛标本，以确定 SPARCL1 的表达情况。用 pIRES2-EGFP-SPARCL1 载体和对照载体转染 HTR8/SVneo 细胞和 JAR 细胞。进行增殖试验和划痕试验。采用实时聚合酶链反应（qPCR）和 Western blot 检测上皮间质转化（EMT）相关分子，包括 MMP2、MMP3、N-钙黏蛋白、E-钙黏蛋白和波形蛋白。采用上述方法检测 HTR8/SVneo 细胞在多次划痕后细胞外信号调节激酶（ERK）磷酸化活性和 AP-1 表达。
流产组的 SPARCL1 mRNA 和蛋白水平明显高于正常妊娠组。转染后，SPARCL1 过表达组与对照组的细胞活力无差异。然而，与对照组相比，SPARCL1 过表达可减少细胞迁移距离和面积，并改变 EMT 相关分子的丰度。SPARCL1 高表达时，ERK 磷酸化活性降低，Fos 和 Jun 表达减少。
滋养层细胞中 SPARCL1 过表达可导致迁移和侵袭能力受限，这可能影响胚胎着床和胎盘发育。它可能参与自然流产的发病机制。
