HGF promotes HTR-8/SVneo cell migration through activation of MAPK/PKA signaling leading to up-regulation of WNT ligands and integrins that target β-catenin

Abstract

Inadequate migration and invasion of the trophoblast cells during embryo implantation is one of the reasons for pregnancy-related complications such as intrauterine growth restriction and preeclampsia. In the present study, relevance of WNT ligands and integrins associated with hepatocyte growth factor (HGF)-mediated migration of HTR-8/SVneo trophoblastic cells has been investigated. Treatment of HTR-8/SVneo cells with HGF led to a dose-dependent increase in their migration. RT-PCR studies revealed a significant increase in the transcripts of WNT4, WNT11, ITGA2, and ITGAV, which was further confirmed at protein level by Western blotting. HGF treatment also led to increased expression of integrin α2β1 and αVβ5 in HTR-8/SVneo cells. Silencing of WNT4, WNT11, ITGA2, and ITGAV by siRNA led to a significant decrease in HGF-mediated migration of cells. Treatment of cells with HGF led to activation of mitogen-activated protein kinases (MAPK) and protein kinase A (PKA) signaling pathways. Inhibition of MAPK/PKA, by selective inhibitors, led to decrease in the expression of above WNT ligands and integrins. Silencing of WNT4/WNT11 led to concomitant decrease in the expression of ITGA2 and ITGAV and vice versa. HGF treatment also led to significant increase in β-catenin expression, a downstream target of both WNT ligands and integrins. Silencing of β-catenin led to decrease in HGF-mediated migration. β-catenin expression was also down-regulated in WNT4/WNT11/ITGA2/ITGAV silenced cells suggesting a possible cross-communication of WNT ligands and integrins via β-catenin. These studies have established the significance of WNT4/WNT11 as well as ITGA2/ITGAV during HGF-mediated migration of HTR-8/SVneo trophoblastic cells.

HGF regulate HTR-8/SVneo trophoblastic cells migration/invasion under hypoxic conditions through increased HIF-1α expression via MAPK and PI3K pathways

“…1 and 2). Significant increase in migration by HGF under normoxic conditions has been previously published by our group (Chaudhary et al 2018). The increase in migration that we observed in HGF treated HTR-8/ SVneo cells might be due to activation of c-Met protein as reported by previous studies in JEG-3 cell line and glioblastoma cell lines under hypoxic conditions (Hayashi et al 2005;Eckerich et al 2007).…”

“…We have previously shown relevance of MAPK and PKA signaling pathways during migration of HTR-8/SVneo cells subsequent to treatment with HGF under normoxia (Chaudhary et al 2018). To investigate, which signaling pathways were activated during HGF-mediated HTR-8/SVneo cells migration/invasion under hypoxic conditions, HTR-8/ SVneo cells were serum starved and subsequently treated with and without HGF for 10, 30 and 60 min under hypoxic conditions.…”

The Involvement of Cell Adhesion Molecules, Tight Junctions, and Gap Junctions in Human Placentation

“…α1β1, α5β1, αvβ3, and αvβ5 promote trophoblast migration and invasion [17][18][19][20][21][22][23]. In addition, α5β1 enhances Fig.…”

Bioinformatic analysis and identification of potential hallmarks in poorly differentiated thyroid cancer

“…Knauf et al [29] found that HGF/Met activation mediates the resistance of mouse anaplastic thyroid cancer to BRAF inhibition, but its mechanism of action is not yet clear. In addition, both EGF and HGF can induce the activation of β-catenin and promote cell migration [30,31], thereby activating the Wnt/β-catenin signaling pathway.. The abnormal activation of Wnt /β-catenin signaling pathway is usually caused by somatic mutations, and its hot-spot mutation genes include: CTNNB1, KRAS, BRAF, NRAS, ERBB2, MET, PIK3CA, JAK1, etc [32].…”