Kisspeptin regulation of genes involved in cell invasion and angiogenesis in first trimester human trophoblast cells.

Ilya Ulasov,Víctor A Francis,Mushi Matjila,Arieh A Katz,Robert P Millar,Aron B Abera

doi:10.1371/journal.pone.0099680

Ilya Ulasov, Víctor A Francis + Show 4 more

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https://doi.org/10.1371/journal.pone.0099680

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Abstract

The precise regulation of extravillous trophoblast invasion of the uterine wall is a key process in successful pregnancies. Kisspeptin (KP) has been shown to inhibit cancer cell metastasis and placental trophoblast cell migration. In this study primary cultures of first trimester human trophoblast cells have been utilized in order to study the regulation of invasion and angiogenesis-related genes by KP. Trophoblast cells were isolated from first trimester placenta and their identity was confirmed by immunostaining for cytokeratin-7. Real-time quantitative RT-PCR demonstrated that primary trophoblast cells express higher levels of GPR54 (KP receptor) and KP mRNA than the trophoblast cell line HTR8Svneo. Furthermore, trophoblast cells also expressed higher GPR54 and KP protein levels. Treating primary trophoblast cells with KP induced ERK1/2 phosphorylation, while co-treating the cells with a KP antagonist almost completely blocked the activation of ERK1/2 and demonstrated that KP through its cognate GPR54 receptor can activate ERK1/2 in trophoblast cells. KP reduced the migratory capability of trophoblast cells in a scratch-migration assay. Real-time quantitative RT-PCR demonstrated that KP treatment reduced the expression of matrix metalloproteinase 1, 2, 3, 7, 9, 10, 14 and VEGF-A, and increased the expression of tissue inhibitors of metalloproteinases 1 and 3. These results suggest that KP can inhibit first trimester trophoblast cells invasion via inhibition of cell migration and down regulation of the metalloproteinase system and VEGF-A.

Highlights

Extravillous trophoblast (EVT) invasion of the maternal uterine wall is a prerequisite for successful placentation and healthy pregnancy
Poor trophoblast invasion is associated with preeclampsia and intrauterine growth restriction (IUGR) [2,3], whereas an excessive invasion leads to placenta accreta or percreta [4]
In order to determine the fraction of trophoblast cells in the primary cultures, cells were stained with anti-cytokeratin-7, which is a positive marker for trophoblast cells and with anti-vimentin which is a marker of mesenchymal cells

Summary

Introduction

Extravillous trophoblast (EVT) invasion of the maternal uterine wall is a prerequisite for successful placentation and healthy pregnancy. During the first trimester of pregnancy, EVTs invade the maternal decidua and the myometrium, remodelling the spiral arteries to ensure an appropriate nutrient and gas exchange between the fetus and the mother [1]. The dysregulation of this process has been shown to cause complications during pregnancy. Trophoblast invasion closely resembles tumour metastasis [5], as trophoblast cells utilize the same molecular mechanisms as cancer cells for their migratory and invasive functions Among these mechanisms, the matrix metalloproteinase (MMP) system is of great importance. The activity of these MMPs can be further regulated by their counterparts, the tissue inhibitors of metalloproteinases (TIMPs) [8]

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