Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Reproductive Biology and Endocrinology
Published in: Reproductive Biology and Endocrinology 1/2009

Open Access 01-12-2009 | Research

Integrin-linked kinase can facilitate syncytialization and hormonal differentiation of the human trophoblast-derived BeWo cell line

Authors: Trina M Butler, Pia A Elustondo, Greg E Hannigan, Daniel J MacPhee

Published in: Reproductive Biology and Endocrinology | Issue 1/2009

Login to get access

Abstract

Background

In the fusion pathway of trophoblast differentiation, stem villous cytotrophoblast cells proliferate and daughter cells differentiate and fuse with existing syncytiotrophoblast to maintain the multi-nucleated layer. Integrin-linked kinase (ILK) is highly expressed in 1st and 2nd trimester villous cytotrophoblast cells, yet barely detectable in syncytiotrophoblast, thus we examined the potential role of ILK in aiding trophoblast fusion.

Methods

The temporal/spatial expression and activity of ILK were determined in BeWo cells undergoing syncytialization by immunoblot and immunofluorescence analyses. BeWo cells were also transfected with pEGFP expression vectors containing wildtype or two mutant ILK cDNA constructs. The incidence of cell fusion in transfected cells grown under syncytialization conditions was then scored by the presence or absence of E-cadherin immunostaining. Beta-hCG expression in transfected cells, a marker of syncytiotrophoblast hormonal differentiation, was also similarly assessed.

Results

ILK catalytic activity increased and ILK began to increasingly localize to BeWo cell nuclei during syncytialization in correlation with increased pAkt and Snail protein expression. Syncytialization was also significantly elevated (p < 0.05) in BeWo cells expressing constitutively active (ca)-ILK vs cells containing empty vector or dn-ILK. Furthermore, cytoplasmic Beta-hCG expression markedly increased (p < 0.05) in cells expressing wt- and ca-ILK.

Conclusion

ILK-facilitated syncytialization is dependent, at least in part, on ILK catalytic activity while hormonal differentiation appears dependent on both ILK-associated protein interactions and catalytic activity. This study demonstrates that ILK plays a novel role in BeWo syncytialization and differentiation, perhaps through an ILK-Akt-Snail pathway, and implicates ILK in the same process in villous cytotrophoblasts in vivo.
Appendix
Available only for authorised users
Literature
1.
Norwitz ER, Schust DJ, Fisher SJ: Implantation and the survival of early pregnancy. New Engl J Med. 2001, 345: 1400-1408. 10.1056/NEJMra000763.CrossRefPubMed
2.
Georgiades P, Ferguson-Smith AC, Burton GJ: Comparative developmental anatomy of the murine and human definitive placentae. Placenta. 2002, 23: 3-19. 10.1053/plac.2001.0738.CrossRefPubMed
3.
Red-Horse K, Zhou Y, Genbacev O, Prakobphol A, Foulk R, McMaster M, Fisher SJ: Trophoblast differentiation during embryo implantation and formation of the maternal-fetal interface. J Clin Invest. 2004, 114: 744-754.PubMedCentralCrossRefPubMed
4.
Bischof P, Irminger-Finger I: The human cytotrophoblastic cell, a mononuclear chameleon. Int J Biochem Cell Biol. 2005, 37: 1-16. 10.1016/j.biocel.2004.05.014.CrossRefPubMed
5.
Kaufmann P, Black S, Huppertz B: Endovascular trophoblast invasion: implications for the pathogenesis of intrauterine growth retardation and preeclampsia. Biol Reprod. 2003, 69: 1-7. 10.1095/biolreprod.102.014977.CrossRefPubMed
6.
Bernischke K, Kaufmann P, Baergen R: Pathology of the human placenta. 2006, New York: Springer Science, 5
7.
Huppertz B, Frank HG, Kingdom JCP, Reister F, Kaufmann P: Villous cytotrophoblast regulation of the syncytial apoptotic cascade in the human placenta. Histochem Cell Biol. 1998, 110: 495-508. 10.1007/s004180050311.CrossRefPubMed
8.
Ellery PM, Cindrova-Davies T, Jauniaux E, Ferguson-Smith AC, Burton GJ: Evidence for transcriptional activity in the syncytiotrophoblast of the human placenta. Placenta. 2009, 30: 329-334. 10.1016/j.placenta.2009.01.002.PubMedCentralCrossRefPubMed
9.
Mori M, Ishikawa G, Luo S-S, Mishima T, Goto T, Robinson JM, Matsubara S, Takeshita T, Kataoka H, Takizawa T: The cytotrophoblast layer of human chorionic villi becomes thinner but maintains its structural integrity during gestation. Biol Reprod. 2007, 76: 164-172. 10.1095/biolreprod.106.056127.CrossRefPubMed
10.
Jones CJ, Harris LK, Whittingham J, Aplin JD, Mayhew TM: A re-appraisal of the morphophenotype and basal lamina coverage of cytotrophoblasts in human term placenta. Placenta. 2008, 29: 215-219. 10.1016/j.placenta.2007.11.004.CrossRefPubMed
11.
Black S, Kadyrov M, Kaufmann P, Ugele B, Emans N, Huppertz B: Syncytial fusion of human trophoblast depends on caspase-8. Cell Death Differ. 2004, 11 (1): 90-98. 10.1038/sj.cdd.4401307.CrossRefPubMed
12.
Huppertz B, Kaufmann P, Kingdom JCP: Trophoblast turnover in health and disease. Fetal Matern Med Rev. 2002, 13: 17-32.CrossRef
13.
Lyden TW, Ng AK, Rote N: Modulation of phosphatidylserine epitope expression by BeWo cells during forskolin treatment. Placenta. 1993, 14: 177-186. 10.1016/S0143-4004(05)80259-0.CrossRefPubMed
14.
Huppertz B, Bartz C, Kokozidou M: Trophoblast fusion: fusogenic proteins, syncytins and ADAMs, and other prerequisites for syncytial fusion. Micron. 2006, 37: 506-517. 10.1016/j.micron.2005.12.011.CrossRef
15.
Frendo J-L, Olivier D, Cheynet V, Blond J-L, Boston O, Vidaud M, Rabreau M, Evain-Brion D, Mallet F: Direct involvement of HERV-W env glycoprotein in human trophoblast cell fusion and differentiation. Mol Cell Biol. 2003, 23: 3566-3574. 10.1128/MCB.23.10.3566-3574.2003.PubMedCentralCrossRefPubMed
16.
Yu C, Shen K, Lin M, Chen P, Lin C, Chang GD, Chen H: GCMa regulates the syncytin-mediated trophoblast fusion. J Biol Chem. 2002, 277: 50062-50068. 10.1074/jbc.M209316200.CrossRefPubMed
17.
Baczyk D, Satkunaratnam A, Nait-Oumesmar B, Huppertz B, Cross JC, Kingdom JC: Complex patterns of GCM1 mRNA and protein in villous and extravillous trophoblast cells of the human placenta. Placenta. 2004, 25: 553-559. 10.1016/j.placenta.2003.12.004.CrossRefPubMed
18.
Daoud G, Rassart E, Masse A, Lafond J: Src family kinases play multiple roles in differentiation of trophoblasts from human term placenta. J Physiol J Physiol. 2006 Mar 15;571(Pt 3):537-53. 2006, 571 (Pt 3): 537-553.
19.
Vargas A, Moreau J, Le Bellego F, Lafond J, Barbeau B: Induction of trophoblast cell fusion by a protein tyrosine phosphatase inhibitor. Placenta. 2008, 29: 70-174. 10.1016/j.placenta.2007.10.012.CrossRef
20.
Boulter E, Van Obberghen-Schilling E: Integrin-linked kinase and its partners: A modular platform regulating cell-matrix adhesion dynamics and cytoskeletal organization. Eur J Cell Biol. 2006, 85: 255-263. 10.1016/j.ejcb.2005.09.002.CrossRefPubMed
21.
Legate KR, Montanez E, Kudlacek O, Fassler R: ILK, PINCH, and parvin: the tIPP of integrin signalling. Nature Mol Cell Bio. 2006, 7: 20-31. 10.1038/nrm1789.CrossRef
22.
Elustondo P, Hannigan GE, Caniggia I, MacPhee DJ: Integrin-Linked Kinase (ILK) is highly expressed in first trimester human chorionic villi and regulates migration of a human cytotrophoblast-derived cell line. Biol Reprod. 2006, 74: 959-968. 10.1095/biolreprod.105.050419.CrossRefPubMed
23.
Yap AS, Crampton MS, Hardin J: Making and breaking contacts: the cellular biology of cadherin regulation. Curr Opin Cell Biol. 2007, 19: 508-514. 10.1016/j.ceb.2007.09.008.PubMedCentralCrossRefPubMed
24.
Coutifaris C, Kao LC, Sehdev HM, Chin U, Babalola GO, Blaschuk OW, Strauss JF: E-cadherin expression during the differentiation of human trophoblasts. Development. 1991, 113 (3): 767-777.PubMed
25.
Getsios S, Chen G, MacCalman C: α-, β-, γ-catenin, and p120ctn, expression during the terminal differentiation and fusion of human mononucleate cytotrophoblast in vitro and in vivo. Mol Reprod Dev. 2001, 59: 168-177. 10.1002/mrd.1019.CrossRefPubMed
26.
Getsios S, MacCalman CD: Cadherin-11 modules the terminal differentiation and fusion of human trophoblastic cells in vitro. Dev Biol. 2003, 257: 41-54. 10.1016/S0012-1606(03)00041-1.CrossRefPubMed
27.
Brown LM, Lacey HA, Baker PN, Crocker IP: E-cadherin in the assessment of aberrant placental cytotrophoblast turnover in pregnancies complicated by pre-eclampsia. Histochem Cell Biol. 2005, 124: 499-506. 10.1007/s00418-005-0051-7.CrossRefPubMed
28.
Hannigan GE, Leung-Hagesteijn C, Fitz-Gibbon L, Coppolino MG, Radeva G, Filmus J, Bell JC, Dedhar S: Regulation of cell adhesion and anchorage-dependent growth by a new β1-integrin-linked protein kinase. Nature. 1996, 379: 91-96. 10.1038/379091a0.CrossRefPubMed
29.
Tan C, Costello P, Sanghera J, Dominguez D, Baulida J, Garcia de Herreros A, Dedhar S: Inhibition of integrin linked kinase (ILK) suppresses β-catenin-Lef/Tcf-dependent transcription and expression of the E-cadherin repressor, snail, in APC -/- human colon carcinoma cells. Oncogene. 2001, 20: 133-140. 10.1038/sj.onc.1204052.CrossRefPubMed
30.
Borges M, Bose P, Frank HG, Kaufmann P, Potgens A: A two colour fluorescence assay for the measurement of syncytial fusion between trophoblast derived cell lines. Placenta. 2003, 24: 959-964. 10.1016/S0143-4004(03)00173-5.CrossRefPubMed
31.
Potgens AJG, Drewlo S, Kokozidou M, Kaufmann P: Syncytin: the major regulator of trophoblast fusion? Recent developments and hypotheses on its action. Hum Reprod Update. 2004, 10 (6): 487-496. 10.1093/humupd/dmh039.CrossRefPubMed
32.
Wice B, Menton D, Geuze H, Schwartz AL: Modulators of cAMP metabolism induce syncytialization formation in vitro. Exp Cell Res. 1990, 186: 306-316. 10.1016/0014-4827(90)90310-7.CrossRefPubMed
33.
34.
Lin L, Xu B, Rote NS: The cellular mechanism by which the human endogenous retrovirus ERV-3 env gene affects proliferation and differentiation in a human placental trophoblast model, BeWo. Placenta. 2000, 21: 73-78. 10.1053/plac.1999.0443.CrossRefPubMed
35.
Kudo Y, Boyd CAR, Sargent IL, Redman CWG, Lee JM, Freeman TC: An analysis using DNA microarray of the time course of gene expression during syncytialization of a human placental cell line (BeWo). Placenta. 2004, 25: 479-488. 10.1016/j.placenta.2003.12.001.CrossRefPubMed
36.
Kudo Y, Boyd CAR: Changes in expression and function of syncytin and its receptor, amino acid transport system B (ASCT2), in human placental choriocarcinoma BeWo cells during syncytialization. Placenta. 2002, 23: 536-541. 10.1053/plac.2002.0839.CrossRefPubMed
37.
Nasiry SA, Spitz B, Hanssens M, Luyten C, Pijnenborg R: Differential effects of inducers of syncytialization and apoptosis on BeWo and JEG-3 choriocarcinoma cells. Hum Reprod. 2006, 21 (1): 193-201. 10.1093/humrep/dei272.CrossRef
38.
Vespa A, Darmon AJ, Turner CE, D'Souza SJA, Dagnino L: Ca2+-dependent localization of integrin-linked kinase to cell junctions in differentiating keratinocytes. J Biol Chem. 2003, 278: 11528-11535. 10.1074/jbc.M208337200.CrossRefPubMed
39.
Vespa A, D'Souza SJ, Dagnino L: A novel role for integrin-linked kinase in epithelial sheet morphogenesis. Mol Biol Cell. 2005, 16: 4084-4095. 10.1091/mbc.E05-02-0087.PubMedCentralCrossRefPubMed
40.
Acconcia F, Barnes CJ, Singh RR, Talukder AH, Kumar R: Phosphorylation-dependent regulation of nuclear localization and functions of integrin-linked kinase. Proc Natl Acad Sci USA. 2007, 104: 6782-6787. 10.1073/pnas.0701999104.PubMedCentralCrossRefPubMed
41.
Das M, Xu B, Lin L, Chakrabarti S, Shivaswamy V, Rote NS: Phosphatidylserine efflux and intercellular fusion in a BeWo model of human villous cytrophoblast. Placenta. 2004, 25: 396-407. 10.1016/j.placenta.2003.11.004.CrossRefPubMed
42.
Daoud G, Amyot M, Rassart E, Masse A, Simoneau L, Lafond J: ERK1/2 and p38 regulate trophoblast differentiation in human term placenta. J Physiol. 2005, 566.2: 409-423. 10.1113/jphysiol.2005.089326.CrossRef
43.
Lynch DK, Ellis CA, Edwards PAW, Hiles ID: Integrin-linked kinase regulates phosphorylation of serine 473 of protein kinase B by an indirect mechanism. Oncogene. 1999, 18: 8024-8032. 10.1038/sj.onc.1203258.CrossRefPubMed
44.
Persad S, Attwell S, Gray V, Delcommenne M, Troussard AA, Sanghera J, Dedhar S: Inhibition of integrin-linked kinase (ILK) suppresses activation of protein kinase B/Akt and induces cell cycle arrest and apoptosis of PTEN-mutant prostate cancer cells. Proc Natl Acad Sci USA. 2000, 97: 3207-3212. 10.1073/pnas.060579697.PubMedCentralCrossRefPubMed
45.
Persad S, Attwell S, Gray V, Mawji N, Deng JT, Leung D, Yan J, Sanghera J, Walsh MP, Dedhar S: Regulation of protein kinase B/Akt-serine 473 phosphorylation by integrin-linked kinase. J Biol Chem. 2001, 276: 27462-27469. 10.1074/jbc.M102940200.CrossRefPubMed
46.
Delcommenne M, Tan C, Gray V, Rue L, Woodgett JR, Dedhar S: Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase. Proc Natl Acad Sci USA. 1998, 95: 11211-11216. 10.1073/pnas.95.19.11211.PubMedCentralCrossRefPubMed
47.
Novak A, Hsu S-C, Leung-Hagesteijn C, Radeva G, Papkoff J, Montesano R, Roskelley C, Grosschedll R, Dedhar S: Proc Natl Acad Sci USA. 1998, 95: 4374-4379. 10.1073/pnas.95.8.4374.PubMedCentralCrossRefPubMed
48.
Yamaji S, Suzuki Y, Sugiyama Y, Koide Y, Yoshida H, Kanamori H, Mohri H, Ohno S, Ishigatsubo Y: A novel integrin-linked kinase-binding protein, affixin, is involved in the early stage of cell-substrate interaction. J Cell Biol. 2001, 153: 1251-1264. 10.1083/jcb.153.6.1251.PubMedCentralCrossRefPubMed
49.
Nikolopoulous SN, Turner CE: Molecular dissection of actopaxin-integrin-linked kinase-paxillin interactions and their role in subcellular localization. J Biol Chem. 2002, 277: 1568-1575. 10.1074/jbc.M108612200.CrossRef
50.
Knerr I, Schubert SW, Wich C, Amann K, Aigner T, Vogler T, Jung R, Dotsch J, Rascher W, Hashelmolhosseini S: Stimulation of GCM1a and syncytin via cAMP mediated PKA signaling in human trophoblastic cells under normoxic and hypoxic conditions. FEBS Lett. 2005, 579: 3991-3998. 10.1016/j.febslet.2005.06.029.CrossRefPubMed
51.
Miller MG, Naruszewicz I, Kumar AS, Ramlal T, Hannigan GE: Integrin-linked kinase is a positive mediator of L6 myoblast differentiation. Biochem Biophys Res Commun. 2003, 310: 796-803. 10.1016/j.bbrc.2003.09.080.CrossRefPubMed
52.
McPhee TR, McDonald PC, Oloumi A, Dedhar S: Integrin-linked kinase regulates E-cadherin expression through PARP-1. Dev Dyn. 2008, 237: 2737-2747. 10.1002/dvdy.21685.CrossRefPubMed
53.
Tan C, Mui A, Dedhar S: Integrin-linked kinase regulates inducible nitric oxide synthase and cyclooxygenase-2 expression in an NF-kappa B-dependent manner. J Biol Chem. 2002, 277: 3109-3116. 10.1074/jbc.M108673200.CrossRefPubMed
54.
Mongroo PS, Johnstone CN, Naruszewicz I, Leung-Hagesteijn C, Sung RK, Carnio L, Rustgi AK, Hannigan GE: Beta-parvin inhibits integrin-linked kinase signaling and is downregulated in breast cancer. Oncogene. 2004, 23: 8959-8970. 10.1038/sj.onc.1208112.CrossRefPubMed
55.
Chen EH, Grote E, Mohler W, Vignery A: Cell-cell fusion. FEBS Lett. 2007, 581: 2181-2193. 10.1016/j.febslet.2007.03.033.CrossRefPubMed
56.
Larsson L-I, Bjerregaard B, Wulf-Andersen L, Talts JF: Syncytin and cancer cell fusions. Directions in Science. 2007, 7: 1193-1197.
Metadata
Title
Integrin-linked kinase can facilitate syncytialization and hormonal differentiation of the human trophoblast-derived BeWo cell line
Authors
Trina M Butler
Pia A Elustondo
Greg E Hannigan
Daniel J MacPhee
Publication date
01-12-2009
Publisher
BioMed Central
Published in
Reproductive Biology and Endocrinology / Issue 1/2009
Electronic ISSN: 1477-7827
DOI
https://doi.org/10.1186/1477-7827-7-51

Other articles of this Issue 1/2009

Reproductive Biology and Endocrinology 1/2009 Go to the issue

Research

The role and molecular mechanism of D-aspartic acid in the release and synthesis of LH and testosterone in humans and rats

Research

Identification and characterization of Ca2+-activated K+ channels in granulosa cells of the human ovary

Research

Endometrial thickness, Caucasian ethnicity, and age predict clinical pregnancy following fresh blastocyst embryo transfer: a retrospective cohort

Research

Predicting lung maturity in preterm rupture of membranes via lamellar bodies count from a vaginal pool: a cohort study

Research

The impact of cocaine and heroin on the placental transfer of methadone

Methodology

Laparoscopic conservative management of ureteral endometriosis: a survey of eighty patients submitted to ureterolysis