Institute of Animal Reproduction and Food Research, The Polish Academy of Sciences, Olsztyn, Poland.
MRC Centre for Reproductive Health, University of Edinburgh, UK.
Placenta. 2019 Feb;77:19-29. doi: 10.1016/j.placenta.2019.01.020. Epub 2019 Jan 24.
The amount of prostaglandin F (PGF) in the uterine lumen increases during the window of implantation in many mammals, including humans. We hypothesized that PGF regulates processes related to human embryo implantation.
The effect of PGF was studied using an in vitro model of human extravillous trophoblast (EVT) cell line (HTR-8/SVneo). Adhesion, proliferation, invasion and migration assays, zymography for metalloproteinases (MMP) activity, and gene/protein expression analyses were applied. Doses of 100 nM and/or 1 μM of PGF and fluprostenol were used. PGF receptor (PTGFR), MMP9 and MMP2 proteins in the human first trimester placenta were localized by immunohistochemistry and immunofluorescence.
This study is the first reporting the expression of PTGFR protein in the first trimester placenta, as well as in HTR-8/SVneo cells. PGF and fluprostenol increased HTR-8/SVneo cell proliferation and adhesion to extracellular matrix protein (P < 0.05). This effect was abolished by mitogen activated protein kinases (MAPK) inhibitor. PGF induced phosphorylation of focal adhesion kinase and MAPK1/3 (P < 0.05). PGF increased mRNA content and protein activity of MMP9, and gene and protein expression of interleukin-6 (P < 0.05). EVT cell migration and invasiveness were stimulated by PGF (P < 0.05). The PGF effect on cell invasion was reduced by inhibitors of MMP2, MMP9 and mTOR. In all experiments, the stimulatory effects of PGF were diminished by using a PTGFR antagonist.
Our findings suggest a significant role for PGF in mechanisms associated with implantation. PGF acting by PTGFR in HTR-8/SVneo cells stimulates their adhesion and proliferation through the MAPK signaling pathway and increases invasiveness inducing MMP proteolytic activity and mTOR signaling.
在许多哺乳动物（包括人类）的着床窗口期，子宫腔中的前列腺素 F（PGF）含量增加。我们假设 PGF 调节与人类胚胎着床相关的过程。
使用人绒毛外滋养层（EVT）细胞系（HTR-8/SVneo）的体外模型研究 PGF 的作用。应用黏附、增殖、侵袭和迁移测定、明胶酶谱法分析金属蛋白酶（MMP）活性，以及基因/蛋白表达分析。使用 100nM 和/或 1μM 的 PGF 和氟前列醇进行处理。通过免疫组织化学和免疫荧光法定位人早孕胎盘中的 PGF 受体（PTGFR）、MMP9 和 MMP2 蛋白。
本研究首次报道了 PTGFR 蛋白在早孕胎盘中以及在 HTR-8/SVneo 细胞中的表达。PGF 和氟前列醇增加了 HTR-8/SVneo 细胞的增殖和对细胞外基质蛋白的黏附（P<0.05）。这种作用被丝裂原活化蛋白激酶（MAPK）抑制剂所阻断。PGF 诱导黏着斑激酶和 MAPK1/3 的磷酸化（P<0.05）。PGF 增加了 MMP9 的 mRNA 含量和蛋白活性，以及白细胞介素-6（IL-6）的基因和蛋白表达（P<0.05）。PGF 刺激 EVT 细胞迁移和侵袭（P<0.05）。MMP2、MMP9 和 mTOR 抑制剂降低了 PGF 对细胞侵袭的作用。在所有实验中，使用 PTGFR 拮抗剂降低了 PGF 的刺激作用。
我们的研究结果表明 PGF 在与着床相关的机制中具有重要作用。PGF 通过 HTR-8/SVneo 细胞中的 PTGFR 作用，通过 MAPK 信号通路刺激其黏附和增殖，并增加侵袭性，诱导 MMP 蛋白水解活性和 mTOR 信号。
