Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan.
Department of Obstetrics and Gynecology, Keiyu Hospital, Yokohama, Japan.
Placenta. 2020 Nov;101:194-203. doi: 10.1016/j.placenta.2020.09.061. Epub 2020 Sep 22.
P2Y14, one of the P2Y purinergic G-protein coupled receptors, is expressed in a variety of cells and tissues. Its ligand, UDP-glucose (UDPG), is released from damaged and stress-stimulated cells and acts as a danger signal via P2Y14. Thus, P2Y14 plays an important role in immunological defense systems. Here, we aimed to elucidate the expression, localization, and role of P2Y14 in human trophoblasts and the placenta.
Human chorionic villus and placental tissues were subjected to immunostaining for P2Y14 protein and an extravillous trophoblast (EVT) marker, HLA-G. We examined the expression of P2Y14 and the effect of UDPG on cell proliferation and invasion in an EVT cell line, HTR-8/SVneo, using an MTS assay and a Transwell assay, respectively. We tested the effect of UDPG on cell invasion in P2Y14-underexpressing HTR-8/SVneo clones established by the lentiviral introduction of shRNA for P2RY14 mRNA.
Immunostaining revealed that P2Y14 was exclusively expressed by EVTs. P2RY14 mRNA and P2Y14 protein were expressed in HTR-8/SVneo cells. UDPG did not affect cell proliferation but it did enhance invasion. Inhibition of P2Y14 and decreasing the expression of P2Y14 suppressed UDPG-mediated invasive activity.
These results showed that EVT selectively expressed P2Y14 and that P2Y14 was positively involved in UDPG-enhanced EVT invasion. It suggests the possible existence of a danger signal-mediated physiological system at the fetomaternal interface where UDPG released from maternal tissues through destruction by EVT invasion may accelerate EVT invasion, allowing EVTs to undergo successful placentation and vascular remodeling.
P2Y14 是 P2Y 嘌呤能 G 蛋白偶联受体家族的一员，在多种细胞和组织中表达。其配体 UDP-葡萄糖（UDPG）从受损和应激刺激的细胞中释放出来，并通过 P2Y14 作为危险信号发挥作用。因此，P2Y14 在免疫防御系统中发挥重要作用。在这里，我们旨在阐明 P2Y14 在人滋养层和胎盘中的表达、定位和作用。
用人绒毛膜绒毛和胎盘组织进行 P2Y14 蛋白和绒毛外滋养层（EVT）标志物 HLA-G 的免疫染色。我们使用 MTS 测定法和 Transwell 测定法分别检查了 EVT 细胞系 HTR-8/SVneo 中 P2Y14 的表达以及 UDPG 对细胞增殖和侵袭的影响。我们使用慢病毒导入针对 P2RY14 mRNA 的 shRNA 建立了 P2Y14 低表达的 HTR-8/SVneo 克隆，测试了 UDPG 对 P2Y14 低表达的 HTR-8/SVneo 克隆细胞侵袭的影响。
免疫染色显示 P2Y14 仅由 EVT 表达。HTR-8/SVneo 细胞中表达 P2RY14 mRNA 和 P2Y14 蛋白。UDPG 不影响细胞增殖，但可增强侵袭。P2Y14 的抑制和 P2Y14 表达的降低抑制了 UDPG 介导的侵袭活性。
这些结果表明 EVT 选择性表达 P2Y14，并且 P2Y14 积极参与 UDPG 增强的 EVT 侵袭。这表明在胎母界面可能存在一种危险信号介导的生理系统，其中 EVT 侵袭通过破坏从母体组织释放的 UDPG 可能加速 EVT 侵袭，允许 EVT 进行成功的胎盘形成和血管重塑。
