Skip to main content
Log in

Insulin-dependent, glucose transporter 1 mediated glucose uptake and tube formation in the human placental first trimester trophoblast cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

During early gestation, hypoxic condition is critically maintained by optimal glucose metabolism and transporter activities. Glucose is readily available energy nutrient required for placentation. However, limited data are available on glucose uptake and its transporters during first trimester placentation processes. To this end, effects of glucose and the roles of glucose transporters (GLUTs) were investigated during hypoxia on trophoblast migration and placental angiogenesis processes using early gestation-derived trophoblast cells, HTR8/SVneo, and first trimester human placental explant tissues. Exogenously added glucose (25 mM) significantly increased tube formation (in vitro angiogenesis) in HTR8/SVneo cells with concomitant activation of AKT-PI3K pathway and increased expression of vascular cell adhesion molecule 1 (VCAM1) compared with those in the presence of 11 mM glucose. Cobalt chloride (CoCl2)-induced hypoxia also significantly increased glucose uptake and GLUT1 expression along with tube formation and migration of HTR8/SVneo cells. During hypoxia, addition of glucose further stimulated HIF1α expression than by hypoxia alone. Cytochalasin B (cyt-B) inhibited the glucose uptake both in the presence of 11 mM and 25 mM glucose. Insulin (1 ng/ml) stimulated GLUT1 expression and tube formation and up-regulated the expression of VEGFR2 in HTR8/SVneo cells. Insulin and glucose-stimulated tube formation was inhibited by cyt-B but had no effect on hypoxia-induced tube formation. Silencing of GLUT1 inhibited the glucose and insulin-stimulated tube formation as well as glucose uptake. However, fatty acid-stimulated tube formation was not affected in GLUT1 knockdown cells. All these data suggest that glucose uptake, glucose-stimulated tube formation, and insulin-stimulated glucose uptake of the first trimester trophoblast cells, HTR8/SVneo, are mediated in part via GLUT1.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Correia-Branco A, Azevedo CF, Araujo JR, Guimaraes JT, Faria A, Keating E, Martel F (2015) Xanthohumol impairs glucose uptake by a human first-trimester extravillous trophoblast cell line (HTR-8/SVneo cells) and impacts the process of placentation. Mol Hum Reprod 21:803–815. https://doi.org/10.1093/molehr/gav043

    Article  CAS  PubMed  Google Scholar 

  2. Frolova A, Flessner L, Chi M, Kim ST, Foyouzi-Yousefi N, Moley KH (2009) Facilitative glucose transporter type 1 is differentially regulated by progesterone and estrogen in murine and human endometrial stromal cells. Endocrinology 150:1512–1520. https://doi.org/10.1210/en.2008-1081

    Article  CAS  PubMed  Google Scholar 

  3. Frolova AI, Moley KH (2011) Quantitative analysis of glucose transporter mRNAs in endometrial stromal cells reveals critical role of GLUT1 in uterine receptivity. Endocrinology 152:2123–2128. https://doi.org/10.1210/en.2010-1266

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Hahn T, Hahn D, Blaschitz A, Korgun ET, Desoye G, Dohr G (2000) Hyperglycaemia-induced subcellular redistribution of GLUT1 glucose transporters in cultured human term placental trophoblast cells. Diabetologia 43:173–180. https://doi.org/10.1007/s001250050026

    Article  CAS  PubMed  Google Scholar 

  5. Illsley NP, Sellers MC, Wright RL (1998) Glycaemic regulation of glucose transporter expression and activity in the human placenta. Placenta 19:517–524

    Article  CAS  PubMed  Google Scholar 

  6. Baumann MU, Deborde S, Illsley NP (2002) Placental glucose transfer and fetal growth. Endocrine 19:13–22. https://doi.org/10.1385/endo:19:1:13

    Article  CAS  PubMed  Google Scholar 

  7. Belkacemi L, Lash GE, Macdonald-Goodfellow SK, Caldwell JD, Graham CH (2005) Inhibition of human trophoblast invasiveness by high glucose concentrations. J Clin Endocrinol Metab 90:4846–4851. https://doi.org/10.1210/jc.2004-2242

    Article  CAS  PubMed  Google Scholar 

  8. Illsley NP (2000) Glucose transporters in the human placenta. Placenta 21:14–22. https://doi.org/10.1053/plac.1999.0448

    Article  CAS  PubMed  Google Scholar 

  9. Sakata M, Kurachi H, Imai T, Tadokoro C, Yamaguchi M, Yoshimoto Y, Oka Y, Miyake A (1995) Increase in human placental glucose transporter-1 during pregnancy. Eur J Endocrinol 132:206–212

    Article  CAS  PubMed  Google Scholar 

  10. Oh S, Kim H, Nam K, Shin I (2017) Glut1 promotes cell proliferation, migration and invasion by regulating epidermal growth factor receptor and integrin signaling in triple-negative breast cancer cells. BMB Reports 50:132–137. https://doi.org/10.5483/BMBRep.2017.50.3.189

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Korgun ET, Celik-Ozenci C, Seval Y, Desoye G, Demir R (2005) Do glucose transporters have other roles in addition to placental glucose transport during early pregnancy? Histochem Cell Biol 123:621–629. https://doi.org/10.1007/s00418-005-0792-3

    Article  CAS  PubMed  Google Scholar 

  12. Heilig C, Brosius F, Siu B, Concepcion L, Mortensen R, Heilig K, Zhu M, Weldon R, Wu G, Conner D (2003) Implications of glucose transporter protein type 1 (GLUT1)-haplodeficiency in embryonic stem cells for their survival in response to hypoxic stress. Am J Pathol 163:1873–1885. https://doi.org/10.1016/s0002-9440(10)63546-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Acosta O, Ramirez VI, Lager S, Gaccioli F, Dudley DJ, Powell TL, Jansson T (2015) Increased glucose and placental GLUT-1 in large infants of obese nondiabetic mothers. Am J Obstet Gynecol 212:227.e1–227.e7. https://doi.org/10.1016/j.ajog.2014.08.009

    Article  CAS  Google Scholar 

  14. Luscher BP, Marini C, Joerger-Messerli MS, Huang X, Hediger MA, Albrecht C, Baumann MU, Surbek DV (2017) Placental glucose transporter (GLUT)-1 is down-regulated in preeclampsia. Placenta 55:94–99. https://doi.org/10.1016/j.placenta.2017.04.023

    Article  CAS  PubMed  Google Scholar 

  15. Chen Y, Zhang Y, Deng Q, Shan N, Peng W, Luo X, Zhang H, Baker PN, Tong C, Qi H (2016) Inhibition of Wnt inhibitory factor 1 under hypoxic condition in human umbilical vein endothelial cells promoted angiogenesis in vitro. Reprod Sci. https://doi.org/10.1177/1933719116638174

    Article  PubMed  PubMed Central  Google Scholar 

  16. Starska K, Forma E, Jozwiak P, Brys M, Lewy-Trenda I, Brzezinska-Blaszczyk E, Krzeslak A (2015) Gene and protein expression of glucose transporter 1 and glucose transporter 3 in human laryngeal cancer-the relationship with regulatory hypoxia-inducible factor-1alpha expression, tumor invasiveness, and patient prognosis. Tumour Biol 36:2309–2321. https://doi.org/10.1007/s13277-014-2838-4

    Article  CAS  PubMed  Google Scholar 

  17. Esterman A, Greco MA, Mitani Y, Finlay TH, Ismail-Beigi F, Dancis J (1997) The effect of hypoxia on human trophoblast in culture: morphology, glucose transport and metabolism. Placenta 18:129–136

    Article  CAS  PubMed  Google Scholar 

  18. Zamudio S, Torricos T, Fik E, Oyala M, Echalar L, Pullockaran J, Tutino E, Martin B, Belliappa S, Balanza E, Illsley NP (2010) Hypoglycemia and the origin of hypoxia-induced reduction in human fetal growth. PLoS ONE 5:e8551. https://doi.org/10.1371/journal.pone.0008551

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Baumann MU, Zamudio S, Illsley NP (2007) Hypoxic upregulation of glucose transporters in BeWo choriocarcinoma cells is mediated by hypoxia-inducible factor-1. Am J Physiol Cell Physiol 293:C477–C485. https://doi.org/10.1152/ajpcell.00075.2007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Hayashi M, Sakata M, Takeda T, Yamamoto T, Okamoto Y, Sawada K, Kimura A, Minekawa R, Tahara M, Tasaka K, Murata Y (2004) Induction of glucose transporter 1 expression through hypoxia-inducible factor 1alpha under hypoxic conditions in trophoblast-derived cells. J Endocrinol 183:145–154. https://doi.org/10.1677/joe.1.05599

    Article  CAS  PubMed  Google Scholar 

  21. Ogura K, Sakata M, Yamaguchi M, Kurachi H, Murata Y (1999) High concentration of glucose decreases glucose transporter-1 expression in mouse placenta in vitro and in vivo. J Endocrinol 160:443–452

    Article  CAS  PubMed  Google Scholar 

  22. Boileau P, Cauzac M, Pereira MA, Girard J, Hauguel-De Mouzon S (2001) Dissociation between insulin-mediated signaling pathways and biological effects in placental cells: role of protein kinase B and MAPK phosphorylation. Endocrinology 142:3974–3979. https://doi.org/10.1210/endo.142.9.8391

    Article  CAS  PubMed  Google Scholar 

  23. Challier JC, Hauguel S, Desmaizieres V (1986) Effect of insulin on glucose uptake and metabolism in the human placenta. J Clin Endocrinol Metab 62:803–807. https://doi.org/10.1210/jcem-62-5-803

    Article  CAS  PubMed  Google Scholar 

  24. Acevedo CG, Marquez JL, Rojas S, Bravo I (2005) Insulin and nitric oxide stimulates glucose transport in human placenta. Life Sci 76:2643–2653. https://doi.org/10.1016/j.lfs.2004.09.039

    Article  CAS  PubMed  Google Scholar 

  25. Brunette MG, Lajeunesse D, Leclerc M, Lafond J (1990) Effect of insulin on D-glucose transport by human placental brush border membranes. Mol Cell Endocrinol 69:59–68

    Article  CAS  PubMed  Google Scholar 

  26. Gordon MC, Zimmerman PD, Landon MB, Gabbe SG, Kniss DA (1995) Insulin and glucose modulate glucose transporter messenger ribonucleic acid expression and glucose uptake in trophoblasts isolated from first-trimester chorionic villi. Am J Obstet Gynecol 173:1089–1097

    Article  CAS  PubMed  Google Scholar 

  27. Kniss DA, Shubert PJ, Zimmerman PD, Landon MB, Gabbe SG (1994) Insulinlike growth factors. Their regulation of glucose and amino acid transport in placental trophoblasts isolated from first-trimester chorionic villi. J Reprod Med 39:249–256

    CAS  PubMed  Google Scholar 

  28. Baumann MU, Schneider H, Malek A, Palta V, Surbek DV, Sager R, Zamudio S, Illsley NP (2014) Regulation of human trophoblast GLUT1 glucose transporter by insulin-like growth factor I (IGF-I). PLoS ONE 9:e106037. https://doi.org/10.1371/journal.pone.0106037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Basak S, Das MK, Srinivas V, Duttaroy AK (2015) The interplay between glucose and fatty acids on tube formation and fatty acid uptake in the first trimester trophoblast cells, HTR8/SVneo. Mol Cell Biochem 401:11–19. https://doi.org/10.1007/s11010-014-2287-9

    Article  CAS  PubMed  Google Scholar 

  30. Chandel NS, Maltepe E, Goldwasser E, Mathieu CE, Simon MC, Schumacker PT (1998) Mitochondrial reactive oxygen species trigger hypoxia-induced transcription. Proc Natl Acad Sci USA 95:11715–11720

    Article  CAS  PubMed  Google Scholar 

  31. Maxwell PH, Wiesener MS, Chang GW, Clifford SC, Vaux EC, Cockman ME, Wykoff CC, Pugh CW, Maher ER, Ratcliffe PJ (1999) The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis. Nature 399:271–275. https://doi.org/10.1038/20459

    Article  CAS  PubMed  Google Scholar 

  32. Yamanaka-Tatematsu M, Nakashima A, Fujita N, Shima T, Yoshimori T, Saito S (2013) Autophagy induced by HIF1α overexpression supports trophoblast invasion by supplying cellular energy. PLoS ONE 8. https://doi.org/10.1371/journal.pone.0076605

  33. Basak S, Duttaroy AK (2012) Leptin induces tube formation in first-trimester extravillous trophoblast cells. Eur J Obstet Gynecol Reprod Biol 164:24–29. https://doi.org/10.1016/j.ejogrb.2012.05.033

    Article  CAS  PubMed  Google Scholar 

  34. Johnsen GM, Basak S, Weedon-Fekjaer MS, Staff AC, Duttaroy AK (2011) Docosahexaenoic acid stimulates tube formation in first trimester trophoblast cells, HTR8/SVneo. Placenta 32:626–632. https://doi.org/10.1016/j.placenta.2011.06.009

    Article  CAS  PubMed  Google Scholar 

  35. Basak S, Sarkar A, Mathapati S, Duttaroy AK (2017) Cellular growth and tube formation of HTR8/SVneo trophoblast: effects of exogenously added fatty acid-binding protein-4 and its inhibitor. Mol Cell Biochem. https://doi.org/10.1007/s11010-017-3095-9

    Article  PubMed  Google Scholar 

  36. Basak S, Duttaroy AK (2013) cis-9,trans-11 conjugated linoleic acid stimulates expression of angiopoietin like-4 in the placental extravillous trophoblast cells. Biochim Biophys Acta 1831:834–843. https://doi.org/10.1016/j.bbalip.2013.01.012

    Article  CAS  PubMed  Google Scholar 

  37. Pandya AD, Das MK, Sarkar A, Vilasagaram S, Basak S, Duttaroy AK (2016) Tube formation in the first trimester placental trophoblast cells: differential effects of angiogenic growth factors and fatty acids. Cell Biol Int 40:652–661. https://doi.org/10.1002/cbin.10601

    Article  CAS  PubMed  Google Scholar 

  38. Barros LF, Yudilevich DL, Jarvis SM, Beaumont N, Baldwin SA (1995) Quantitation and immunolocalization of glucose transporters in the human placenta. Placenta 16:623–633

    Article  CAS  PubMed  Google Scholar 

  39. Gunnink LK, Alabi OD, Kuiper BD, Gunnink SM, Schuiteman SJ, Strohbehn LE, Hamilton KE, Wrobel KE, Louters LL (2016) Curcumin directly inhibits the transport activity of GLUT1. Biochimie. https://doi.org/10.1016/j.biochi.2016.03.014

    Article  PubMed  PubMed Central  Google Scholar 

  40. Sage JM, Cura AJ, Lloyd KP, Carruthers A (2015) Caffeine inhibits glucose transport by binding at the GLUT1 nucleotide-binding site. Am J Physiol Cell Physiol 308:C827–C834. https://doi.org/10.1152/ajpcell.00001.2015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Beckert S, Farrahi F, Aslam RS, Scheuenstuhl H, Konigsrainer A, Hussain MZ, Hunt TK (2006) Lactate stimulates endothelial cell migration. Wound Repair Regen 14:321–324. https://doi.org/10.1111/j.1743-6109.2006.00127.x

    Article  PubMed  Google Scholar 

  42. Vallee A, Guillevin R, Vallee JN (2017) Vasculogenesis and angiogenesis initiation under normoxic conditions through Wnt/beta-catenin pathway in gliomas. Rev Neurosci. https://doi.org/10.1515/revneuro-2017-0032

    Article  Google Scholar 

  43. Desoye G, Hartmann M, Blaschitz A, Dohr G, Hahn T, Kohnen G, Kaufmann P (1994) Insulin receptors in syncytiotrophoblast and fetal endothelium of human placenta. Immunohistochemical evidence for developmental changes in distribution pattern. Histochemistry 101:277–285

    Article  CAS  PubMed  Google Scholar 

  44. Carayannopoulos MO, Chi MM, Cui Y, Pingsterhaus JM, McKnight RA, Mueckler M, Devaskar SU, Moley KH (2000) GLUT8 is a glucose transporter responsible for insulin-stimulated glucose uptake in the blastocyst. Proc Natl Acad Sci USA 97:7313–7318

    Article  CAS  PubMed  Google Scholar 

  45. Barthel A, Okino ST, Liao J, Nakatani K, Li J, Whitlock JP Jr, Roth RA (1999) Regulation of GLUT1 gene transcription by the serine/threonine kinase Akt1. J Biol Chem 274:20281–20286

    Article  CAS  PubMed  Google Scholar 

  46. Sweeney G, Somwar R, Ramlal T, Volchuk A, Ueyama A, Klip A (1999) An inhibitor of p38 mitogen-activated protein kinase prevents insulin-stimulated glucose transport but not glucose transporter translocation in 3T3-L1 adipocytes and L6 myotubes. J Biol Chem 274:10071–10078

    Article  CAS  PubMed  Google Scholar 

  47. Marini M, Vichi D, Toscano A, Thyrion GDZ, Bonaccini L, Parretti E, Gheri G, Pacini A, Sgambati E (2008) Effect of impaired glucose tolerance during pregnancy on the expression of VEGF receptors in human placenta. Reprod Fertil Dev. https://doi.org/10.1071/RD08032

    Article  PubMed  Google Scholar 

  48. Lassance L, Miedl H, Absenger M, Diaz-Perez F, Lang U, Desoye G, Hiden U (2013) Hyperinsulinemia stimulates angiogenesis of human fetoplacental endothelial cells: a possible role of insulin in placental hypervascularization in diabetes mellitus. J Clin Endocrinol Metab 98:E1438–E1447. https://doi.org/10.1210/jc.2013-1210

    Article  CAS  PubMed  Google Scholar 

  49. Liu Y, Petreaca M, Martins-Green M (2009) Cell and molecular mechanisms of insulin-induced angiogenesis. J Cell Mol Med 13:4492–4504. https://doi.org/10.1111/j.1582-4934.2008.00555.x

    Article  CAS  PubMed  Google Scholar 

  50. Ericsson A, Hamark B, Powell TL, Jansson T (2005) Glucose transporter isoform 4 is expressed in the syncytiotrophoblast of first trimester human placenta. Hum Reprod 20:521–530. https://doi.org/10.1093/humrep/deh596

    Article  CAS  PubMed  Google Scholar 

  51. Vollers SS, Carruthers A (2012) Sequence determinants of GLUT1-mediated accelerated-exchange transport: analysis by homology-scanning mutagenesis. J Biol Chem 287:42533–42544. https://doi.org/10.1074/jbc.M112.369587

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Mayer A, Schmidt M, Seeger A, Serras AF, Vaupel P, Schmidberger H (2014) GLUT-1 expression is largely unrelated to both hypoxia and the Warburg phenotype in squamous cell carcinomas of the vulva. BMC Cancer 14:760. https://doi.org/10.1186/1471-2407-14-760

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This collaborative work was initiated and started at the University of Oslo, Norway while SB worked as a visiting scientist (Boyscast Fellowship).V.S was supported by the fellowship of Council of Scientific & Industrial Research (CSIR), India. This study was supported by a grant (BT/PR6946/MED/97/89/2012) received from the Department of Biotechnology, Government of India.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sanjay Basak.

Ethics declarations

Conflict of interest

The authors have no conflict of interests to disclose.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 18 KB)

Supplementary Fig. 1

Expression of angiogenic growth factors in response to glucose pre-stimulation of HTR8/SVneo cells. Cells were pre-stimulated with glucose (5.5mM, 11mM and 25mM) for 24h. mRNA expression of each gene was analyzed using quantitative real-time RT-PCR and normalized the expression to the endogenous control TBP. Fold change of expression was calculated according to the ΔΔCt method. Data are presented as fold expression of control (mean± SEM, n=9) those were analyzed by Student’s t test, * p<0.05. Supplementary material 2 (TIF 28711 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Basak, S., Vilasagaram, S., Naidu, K. et al. Insulin-dependent, glucose transporter 1 mediated glucose uptake and tube formation in the human placental first trimester trophoblast cells. Mol Cell Biochem 451, 91–106 (2019). https://doi.org/10.1007/s11010-018-3396-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-018-3396-7

Keywords

Navigation