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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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Abstract

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.

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References

  • Anacker J, Segerer SE, Hagemann C, Feix S, Kapp M, Bausch R, Kämmerer U (2011) Human decidua and invasive trophoblasts are rich sources of nearly all human matrix metalloproteinases. Mol Hum Reprod 17:637–652

    CAS  PubMed  Google Scholar 

  • Aplin JD (1991) Implantation, trophoblast differentiation and haemochorial placentation: mechanistic evidence in vivo and in vitro. J Cell Sci 99:681–692

    PubMed  Google Scholar 

  • Burton GJ, Jauniaux E, Watson AL (1999) Maternal arterial connections to the placental intervillous space during the first trimester of human pregnancy: the Boyd collection revisited. Am J Obstet Gynecol 181:718–724

    CAS  PubMed  Google Scholar 

  • Burton GJ, Hempstock J, Jauniaux E (2003) Oxygen, early embryonic metabolism and free radical-mediated embryopathies. Reprod BioMed Online 6:84–96

    PubMed  Google Scholar 

  • Caniggia I, Winter JL (2002) Adriana and Luisa Castellucci award lecture 2001. Hypoxia inducible factor-1: oxygen regulation of trophoblast differentiation in normal and preeclamptic pregnancies-a review. Placenta 23:S47–S57

    PubMed  Google Scholar 

  • Caniggia I, Mostachfi H, Winter J, Gassmann M, Lye SJ (2000) Hypoxia-inducible factor-1 mediates the biological effects of oxygen on human trophoblast differentiation through TGFβ3. J Clin Invest 105:577–587

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cartwright JE, Tse WK, Whitley GS (2002) Hepatocyte growth factor induced human trophoblast motility involves phosphatidylinositol-3-kinase, mitogen-activated protein kinase, and inducible nitric oxide synthase. Exp Cell Res 279:219–226

    CAS  PubMed  Google Scholar 

  • Chaudhary P, Malhotra SS, Babu GS, Sobti RC, Gupta SK (2018) 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. Mol Cell Biochem. https://doi.org/10.1007/s11010-018-3428-3

    PubMed  Google Scholar 

  • Cockle JV, Gopichandran N, Walker JJ, Levene MI, Orsi NM (2007) Matrix metalloproteinases and their tissue inhibitors in preterm perinatal complications. Reprod Sci 14:629–645

    CAS  PubMed  Google Scholar 

  • Coppock H, White A, Aplin JD, Westwood M (2004) Matrix metalloprotease-3 and -9 proteolyze insulin-like growth factor-binding protein-1. Biol Reprod 71:438–443

    CAS  PubMed  Google Scholar 

  • Deng CL, Ling ST, Liu XQ, Zhao YJ, Lv YF (2015) Decreased expression of matrix metalloproteinase-1 in the maternal umbilical serum, trophoblasts and decidua leads to preeclampsia. Exp Ther Med 9:992–998

    CAS  PubMed  PubMed Central  Google Scholar 

  • Dunwoodie SL (2009) The role of hypoxia in development of the mammalian embryo. Dev Cell 17:755–773

    CAS  PubMed  Google Scholar 

  • Eckerich C, Zapf S, Fillbrandt R, Loges S, Westphal M, Lamszus K (2007) Hypoxia can induce c-met expression in glioma cells and enhance SF/HGF-induced cell migration. Int J Cancer 121:276–283

    CAS  PubMed  Google Scholar 

  • Ferrer-Agüero JM, Ferreres J, Mora ML, Lubillo S, Sánchez M, Barrios Y, Sierra A, Páramo JA (2009) Matrix metalloproteinase-9, −10, and tissue inhibitor of matrix metalloproteinases-1 blood levels as biomarkers of severity and mortality in sepsis. Crit Care 13:R158

    PubMed  PubMed Central  Google Scholar 

  • Fisher SJ (2015) Why is placentation abnormal in preeclampsia? Am J Obstet Gynecol 213:115–122

    Google Scholar 

  • Fortunato SJ, Menon R, Lombardi SJ (1999) MMP/TIMP imbalance in amniotic fluid during PROM: an indirect support for endogenous pathway to membrane rupture. J Perinat Med 27:362–368

    CAS  PubMed  Google Scholar 

  • Genbacev O, Joslin R, Damsky CH, Polliotti BM, Fisher SJ (1996) Hypoxia alters early gestation human cytotrophoblast differentiation/invasion in vitro and models the placental defects that occur in preeclampsia. J Clin Invest 97:540–550

    CAS  PubMed  PubMed Central  Google Scholar 

  • Genbacev O, Zhou Y, Ludlow JW, Fisher SJ (1997) Regulation of human placental development by oxygen tension. Science 277:1669–1672

    CAS  PubMed  Google Scholar 

  • Graham CH, Fitzpatrick TE, McCrae KR (1998) Hypoxia stimulates urokinase receptor expression through a heme protein-dependent pathway. Blood 91:3300–3307

    CAS  PubMed  Google Scholar 

  • Ha JG, Li L, Lee DH, Na SH, Ha KS, Kim YM, Ko JH, Hwang JY (2015) The effects of heme oxygenase by-products on the proliferation and invasion of HUVECs, HTR-8/SVneo cells, 3A (tPA 30-1) cells and HESCs under varying oxygen concentrations. Reprod Sci 22:1530–1538

    CAS  PubMed  Google Scholar 

  • Hayashi M, Sakata M, Takeda T, Tahara M, Yamamoto T, Okamoto Y, Minekawa R, Isobe A, Ohmichi M, Tasaka K, Murata YT (2005) Up-regulation of c-met protooncogene product expression through hypoxia-inducible factor-1alpha is involved in trophoblast invasion under low-oxygen tension. Endocrinology 146:4682–4689

    CAS  PubMed  Google Scholar 

  • Huppertz B, Kertschanska S, Demir AY, Frank HG, Kaufmann P (1998) Immunohistochemistry of matrix metalloproteinases (MMP), their substrates, and their inhibitors (TIMP) during trophoblast invasion in the human placenta. Cell Tissue Res 291:133–148

    CAS  PubMed  Google Scholar 

  • Husslein H, Haider S, Meinhardt G, Prast J, Sonderegger S, Knöfler M (2009) Expression, regulation and functional characterization of matrix metalloproteinase-3 of human trophoblast. Placenta 30:284–291

    CAS  PubMed  PubMed Central  Google Scholar 

  • Jauniaux E, Hempstock J, Greenwold N, Burton GJ (2003) Trophoblastic oxidative stress in relation to temporal and regional differences in maternal placental blood flow in normal and abnormal early pregnancies. Am J Pathol 162:115–125

    PubMed  PubMed Central  Google Scholar 

  • Kauma SW, Bae-Jump V, Walsh SW (1999) Hepatocyte growth factor stimulates trophoblast invasion: a potential mechanism for abnormal placentation in preeclampsia. J Clin Endocrinol Metab 84:4092–4096

    CAS  PubMed  Google Scholar 

  • Kobara H, Miyamoto T, Suzuki A, Asaka R, Yamada Y, Ishikawa K, Kikuchi N, Ohira S, Shiozawa T (2013) Lipocalin2 enhances the matrix metalloproteinase-9 activity and invasion of extravillous trophoblasts under hypoxia. Placenta 34:1036–1043

    CAS  PubMed  Google Scholar 

  • Lash GE, Hornbuckle J, Brunt A, Kirkley M, Searle RF, Robson SC, Bulmer JN (2007) Effect of low oxygen concentrations on trophoblast-like cell line invasion. Placenta 28:390–398

    CAS  PubMed  Google Scholar 

  • Lian IA, Toft JH, Olsen GD, Langaas M, Bjørge L, Eide IP, Børdahl PE, Austgulen R (2010) Matrix metalloproteinase1 in pre-eclampsia and fetal growth restriction: reduced gene expression in decidual tissue and protein expression in extravillous trophoblasts. Placenta 31:615–620

    CAS  PubMed  Google Scholar 

  • Liu HY, Jia XQ, Gao LX, Ma YY (2012) Hepatocyte growth factor regulates HLX1 gene expression to modulate HTR-8/SVneo trophoblast cells. Reprod Biol Endocrinol 10:83

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liu L, Wang Y, Shen C, He J, Liu X, Ding Y, Gao R, Chen X (2016) Benzo(a) pyrene inhibits migration and invasion of extravillous trophoblast HTR-8/SVneo cells via activation of the ERK and JNK pathway. J Appl Toxicol 36:946–955

    PubMed  Google Scholar 

  • Luo J, Qiao F, Yin X (2011) Hypoxia induces FGF2 production by vascular endothelial cells and alters MMP9 and TIMP1 expression in extravillous trophoblasts and their invasiveness in a co-cultured model. J Reprod Dev 57:84–91

    CAS  PubMed  Google Scholar 

  • Malik A, Pal R, Gupta SK (2017) Interdependence of JAK-STAT and MAPK signaling pathways during EGF-mediated HTR-8/SVneo cell invasion. PLoS One 12:e0178269

    PubMed  PubMed Central  Google Scholar 

  • Maquoi E, Polette M, Nawrocki B, Bischof P, Noel A, Pintiaux A, Santavicca M, Schaaps JP, Pijnenborg R, Birembaut P, Foidart JM (1997) Expression of stromelysin-3 in the human placenta and placental bed. Placenta 18:277–285

    CAS  PubMed  Google Scholar 

  • Matsushita A, Götze T, Korc M (2007) Hepatocyte growth factor-mediated cell invasion in pancreatic cancer cells is dependent on neuropilin-1. Cancer Res 67:10309–10316

    CAS  PubMed  Google Scholar 

  • Meade ES, Ma YY, Guller S (2007) Role of hypoxia-inducible transcription factors1α and 2α in the regulation of plasminogen activator inhibitor-1 expression in a human trophoblast cell line. Placenta 28:1012–1019

    CAS  PubMed  PubMed Central  Google Scholar 

  • Minet E, Arnould T, Michel G, Roland I, Mottet D, Raes M, Remacle J, Michiels C (2000) ERK activation upon hypoxia: involvement in HIF-1 activation. FEBS Lett 468:53–58

    CAS  PubMed  Google Scholar 

  • Nishi H, Nakada T, Hokamura M, Osakabe Y, Itokazu O, Huang LE, Isaka K (2004) Hypoxia-inducible factor-1 transactivates transforming growth factor-beta3 in trophoblast. Endocrinology 145:4113–4118

    CAS  PubMed  Google Scholar 

  • Onogi A, Naruse K, Sado T, Tsunemi T, Shigetomi H, Noguchi T, Yamada Y, Akasaki M, Oi H, Kobayashi H (2011) Hypoxia inhibits invasion of extravillous trophoblast cells through reduction of matrix metalloproteinase (MMP)-2 activation in the early first trimester of human pregnancy. Placenta 32:665–670

    CAS  PubMed  Google Scholar 

  • Poston L, Raijmakers MT (2004) Trophoblast oxidative stress, antioxidants and pregnancy outcome-a review. Placenta 25:S72–S78

    CAS  PubMed  Google Scholar 

  • Pringle KG, Kind KL, Sferruzzi-Perri AN, Thompson JG, Roberts CT (2010) Beyond oxygen: complex regulation and activity of hypoxia inducible factors in pregnancy. Hum Reprod Update 16:415–431

    CAS  PubMed  Google Scholar 

  • Radtke S, Milanovic M, Rosse C, DeRycker M, Lachmann S, Hibbert A, Kermorgant S, Parker PJ (2013) ERK2 but not ERK1 mediates HGF-induced motility in non-small cell lung carcinoma cell lines. J Cell Sci 126:2381–2391

    CAS  PubMed  Google Scholar 

  • Reister F, Kingdom JC, Ruck P, Marzusch K, Heyl W, Pauer U, Kaufmann P, Rath W, Huppertz B (2006) Altered protease expression by peri-arterial trophoblast cells in severe early-onset preeclampsia with IUGR. J Perinat Med 34:272–279

    CAS  PubMed  Google Scholar 

  • Rodesch F, Simon P, Donner C, Jauniaux E (1992) Oxygen measurements in endometrial and trophoblastic tissues during early pregnancy. Obstet Gynecol 80:283–285

    CAS  PubMed  Google Scholar 

  • Rooth G, Sjosted S, Caligara F (1961) Hydrogen concentration, carbon dioxide tension and acid base balance in blood of human umbilical cord and intervillous space of placenta. Arch Dis Child 36:278–285

    CAS  PubMed  PubMed Central  Google Scholar 

  • Semenza GL (2003) Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3:721–732

    CAS  PubMed  Google Scholar 

  • Semenza GL, Wang GL (1992) A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcriptional activation. Mol Cell Biol 12:5447–5454

    CAS  PubMed  PubMed Central  Google Scholar 

  • Somerset DA, Li XF, Afford S, Strain AJ, Ahmed A, Sangha RK, Whittle MJ, Kilby MD (1998) Ontogeny of hepatocyte growth factor (HGF) and its receptor (c-met) in human placenta: reduced HGF expression in intrauterine growth restriction. Am J Pathol 153:1139–1147

    CAS  PubMed  PubMed Central  Google Scholar 

  • Staun-Ram E, Goldman S, Gabarin D, Shalev E (2004) Expression and importance of matrix metalloproteinase 2 and 9 (MMP-2 and -9) in human trophoblast invasion. Reprod Biol Endocrinol 4:59

    Google Scholar 

  • Sundrani D, Narang A, Mehendale S, Joshi S, Chavan-Gautam P (2017) Investigating the expression of MMPs and TIMPs in preterm placenta and role of CpG methylation in regulating MMP-9 expression. IUBMB Life 69:985–993

    CAS  PubMed  Google Scholar 

  • Tacchini L, Dansi P, Matteucci E, Desiderio MA (2001) Hepatocyte growth factor signaling stimulates hypoxia inducible factor-1 (HIF-1) activity in HepG2 hepatoma cells. Carcinogenesis 22:1363–1371

    CAS  PubMed  Google Scholar 

  • Tacchini L, DePonti C, Matteucci E, Follis R, Desiderio MA (2004) Hepatocyte growth factor-activated NF-kB regulates HIF-1 activity and ODC expression, implicated in survival, differently in different carcinoma cell lines. Carcinogenesis 25:2089–2100

    CAS  PubMed  Google Scholar 

  • Tamaru S, Mizuno Y, Tochigi H, Kajihara T, Okazaki Y, Okagaki R, Kamei Y, Ishihara O, Itakura A (2015) MicroRNA-135b suppresses extravillous trophoblast-derived HTR-8/SVneo cell invasion by directly down regulating CXCL12 under low oxygen conditions. Biochem Biophys Res Commun 461:421–426

    CAS  PubMed  Google Scholar 

  • Van Patot MC, Ebensperger G, Gassmann M, Llanos AJ (2012) The hypoxic placenta. High Alt Med Biol 13:176–184

    PubMed  Google Scholar 

  • Visse R, Nagase H (2003) Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function and biochemistry. Circ Res 92:827–839

    CAS  PubMed  Google Scholar 

  • Wang L, Yu Y, Guan H, Liu T, Qiao C (2015) 67-kDa laminin receptor contributes to hypoxia-induced migration and invasion of trophoblast-like cells by mediating matrix metalloproteinase-9. Clin Exp Pharmacol Physiol 42:549–558

    CAS  PubMed  Google Scholar 

  • Weiss A, Goldman S, Shalev E (2007) The matrix metalloproteinases (MMPs) in the decidua and fetal membranes. Front Biosci 12(1):649–659

    CAS  PubMed  Google Scholar 

  • Wolf HK, Zarnegar R, Oliver L, Michalopoulos GK (1991) Hepatocyte growth factor in human placenta and trophoblastic disease. Am J Pathol 138:1035–1043

    CAS  PubMed  PubMed Central  Google Scholar 

  • Xu L, Pathak PS, Fukumura D (2004) Hypoxia-induced activation of p38 mitogen-activated protein kinase and phosphatidylinositol 3′-kinase signaling pathways contributes to expression of interleukin 8 in human ovarian carcinoma cells. Clin Cancer Res 10:701–707

    CAS  PubMed  Google Scholar 

  • Zhou Y, Genbacev O, Damsky CH, Fisher SJ (1998) Oxygen regulates human cytotrophoblast differentiation and invasion: implications for endovascular invasion in normal pregnancy and in pre-eclampsia. J Reprod Immunol 39:197–192

    CAS  PubMed  Google Scholar 

  • Zhu JY, Pang ZJ, Yu YH (2012) Regulation of trophoblast invasion: the role of matrix metalloproteinases. Rev Obstet Gynecol 5:e137–e143

    PubMed  PubMed Central  Google Scholar 

  • Zhu J, Wang K, Li T, Chen J, Xie D, Chang X, Yao J, Wu J, Zhou Q, Jia Y, Duan T (2017) Hypoxia-induced TET1 facilitates trophoblast cell migration and invasion through HIF1α signaling pathway. Sci Rep 7:8077

    PubMed  PubMed Central  Google Scholar 

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Acknowledgements

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.

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  1. Reproductive Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110 067, India

    Piyush Chaudhary & Satish Kumar Gupta

  2. Department of Biotechnology, Babasaheb Bhimrao Ambedkar University, Vidya Vihar, Lucknow, 226 025, India

    Piyush Chaudhary & Gosipatala Sunil Babu

  3. Department of Biotechnology, Panjab University, Sector-14, Chandigarh, 160 014, India

    Ranbir Chander Sobti

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  1. Piyush Chaudhary
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  2. Gosipatala Sunil Babu
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  4. Satish Kumar Gupta
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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.

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Correspondence to Satish Kumar Gupta.

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Chaudhary, P., Babu, G.S., Sobti, R.C. et al. HGF regulate HTR-8/SVneo trophoblastic cells migration/invasion under hypoxic conditions through increased HIF-1α expression via MAPK and PI3K pathways. J. Cell Commun. Signal. 13, 503–521 (2019). https://doi.org/10.1007/s12079-019-00505-x

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