HGF regulate HTR-8/SVneo trophoblastic cells migration/invasion under hypoxic conditions through increased HIF-1α expression via MAPK and PI3K pathways
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Hepatocyte growth factor (HGF) is reported to be down-regulated in pregnancy complications like intrauterine growth retardation and preeclampsia, which are associated with abnormal trophoblast migration/invasion. In this study, role of HGF and associated signaling pathways has been investigated in HTR-8/SVneo trophoblastic cells migration/invasion under normoxia (20% O2) and hypoxia (2% O2). HTR-8/SVneo cells exposed to hypoxia showed increase in migration and invasion as compared to cells incubated under normoxic conditions. The migration/invasion under both normoxic and hypoxic conditions was further enhanced after treatment with HGF. Subsequent to treatment with HGF, a significant increase in expression of MMP2 & MMP3 under normoxia and MMP1 & MMP9 under hypoxia was observed. Treatment of HTR-8/SVneo cells with HGF under hypoxia also led to decrease in TIMP1. Treatment of the cells with HGF led to activation of mitogen activated protein kinases (MAPK) and phosphatidylinositol 3-kinase (PI3K) signaling pathways. Inhibition of MAPK by U0126 and PI3K by LY294002 led to concomitant decrease in the HGF-mediated migration/invasion of HTR-8/SVneo cells. HGF treatment under hypoxia also led to a significant increase in hypoxia inducible factor (HIF-1α) expression. Additionally, inhibition of HIF-1α by siRNA led to decrease in HGF-mediated migration of HTR-8/SVneo cells under hypoxic conditions. Inhibition of HGF activated MAPK and PI3K signaling led to reduction in HIF-1α expression under hypoxia. In conclusion, HGF facilitates HTR-8/SVneo cell migration/invasion by activation of MAPK/PI3K signaling pathways and increased expression of MMPs. HIF-1α has a role in HGF-mediated increase in migration under hypoxic conditions.
KeywordsHGF HIF-1α Invasion Migration MAPK MMPs PI3K TIMPs
This work was supported by Department of Health Research, Government of India under grant (GIA/28/2014); Department of Biotechnology, Government of India under grant (BT/PR12312/MED/30/1424/2014); J. C. Bose National Fellowship by Science and Engineering Research Board, Department of Science and Technology, Government of India to SKG under grant (SB/S2/JCB-040/2015). SKG would like to acknowledge National Institute of Immunology, New Delhi, India for additional financial support. The funding bodies were not involved in the design of the study, data analysis or writing the manuscript.
PC, SKG and GSB were involved in project conception and experimental design. PC carried out the experiments. PC, SKG, GSB and RCS interpreted the data thus obtained and were involved in writing the manuscript.
Compliance with ethical standards
The authors declare that they have no competing interests.
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