Down-regulation of the transcription factor snail in the placentas of patients with preeclampsia and in a rat model of preeclampsia

Reproductive Biology and Endocrinology

Terrence Horrigan²  Deepak Malhotra¹  Joseph I Shapiro¹  Nauman Khurshid²  Sleiman Smaili²  Cara Gatto-Weis³  Larisa Fedorova¹ 

[1] Department of Medicine, University of Toledo School of Medicine, Toledo, OH 43614, USA;Department of Obstetrics and Gynecology, University of Toledo School of Medicine, Toledo, OH 43614, USA;Department of Pathology, University of Toledo School of Medicine, Toledo, OH 43614, USA

关键词: E-cadherin;    Trophoblast;    Snail;    Placenta;    Preeclampsia;   

Others  :  1150911
DOI  :  10.1186/1477-7827-10-15

 received in 2011-09-30, accepted in 2012-02-23,  发布年份 2012

【 摘 要 】

Background

Placental malfunction in preeclampsia is believed to be a consequence of aberrant differentiation of trophoblast lineages and changes in utero-placental oxygenation. The transcription factor Snail, a master regulator molecule of epithelial-mesenchymal transition in embryonic development and in cancer, is shown to be involved in trophoblast differentiation as well. Moreover, Snail can be controlled by oxidative stress and hypoxia. Therefore, we examined the expression of Snail and its downstream target, e-cadherin, in human normal term, preterm and preeclamptic placentas, and in pregnant rats that developed preeclampsia-like symptoms in the response to a 20-fold increase in sodium intake.

Methods

Western blotting analysis was used for comparative expression of Snail and e- cadherin in total protein extracts. Placental cells expressing Snail and e-cadherin were identified by immunohistochemical double-labeling technique.

Results

The levels of Snail protein were decreased in human preeclamptic placentas by 30% (p < 0.01) compared to normal term, and in the rat model by 40% (p < 0.001) compared to control placentas. In preterm placentas, the levels of Snail expression varied, yet there was a strong trend toward statistical significance between preterm and preeclamptic placentas. In humans, e-cadherin protein level was 30% higher in preeclamptic (p < 0.05) placentas and similarly, but not significantly (p = 0.1), high in the preterm placentas compared to normal term. In the rat model of preeclampsia, e-cadherin was increased by 60% (p < 0.01). Immunohistochemical examination of human placentas demonstrated Snail-positive staining in the nuclei of the villous trophoblasts and mesenchymal cells and in the invasive trophoblasts of the decidua. In the rat placenta, the majority of Snail positive cells were spongiotrophoblasts of the junctional zone, while in the labyrinth, Snail-positive sinusoidal giant trophoblasts cells were found in some focal areas located close to the junctional zone.

Conclusion

We demonstrated that human preeclampsia and the salt-induced rat model of preeclampsia are associated with the reduced levels of Snail protein in placenta. Down-regulation of the transcription factor Snail in placental progenitor cell lineages, either by intrinsic defects and/or by extrinsic and maternal factors, may affect normal placenta development and function and thus contribute to the pathology of preeclampsia.

【 授权许可】

   
2012 Fedorova et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Steegers EA, von Dadelszen P, Duvekot JJ, Pijnenborg R: Pre-eclampsia. Lancet 2010, 376(9741):631-644.
• [2]Benirschke K, Kaufmann P, Baergen R: Pathology of the human placenta. 5th edition. New York: Springer; 2006.
• [3]Huppertz B: Placental origins of preeclampsia: challenging the current hypothesis. Hypertension 2008, 51(4):970-975.
• [4]Burton GJ, Jones CJ: Syncytial knots, sprouts, apoptosis, and trophoblast deportation from the human placenta. Taiwan J Obstet Gynecol 2009, 48(1):28-37.
• [5]Huppertz B: IFPA Award in Placentology Lecture: biology of the placental syncytiotrophoblast-myths and facts. Placenta 2010, 31(Suppl):S75-S81.
• [6]Cindrova-Davies T: Gabor Than Award Lecture 2008: pre-eclampsia--from placental oxidative stress to maternal endothelial dysfunction. Placenta 2009, 30(Suppl A):S55-S65.
• [7]Heazell AE, Lacey HA, Jones CJ, Huppertz B, Baker PN, Crocker IP: Effects of oxygen on cell turnover and expression of regulators of apoptosis in human placental trophoblast. Placenta 2008, 29(2):175-186.
• [8]Zhou Y, Damsky CH, Fisher SJ: Preeclampsia is associated with failure of human cytotrophoblasts to mimic a vascular adhesion phenotype. One cause of defective endovascular invasion in this syndrome? J Clin Invest 1997, 99(9):2152-2164.
• [9]Thiery JP, Acloque H, Huang RY, Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell 2009, 139(5):871-890.
• [10]Rosario GX, Ain R, Konno T, Soares MJ: Intrauterine fate of invasive trophoblast cells. Placenta 2009, 30(5):457-463.
• [11]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(6):499-506.
• [12]Zhou Y, Fisher SJ, Janatpour M, Genbacev O, Dejana E, Wheelock M, Damsky CH: Human cytotrophoblasts adopt a vascular phenotype as they differentiate. A strategy for successful endovascular invasion? J Clin Invest 1997, 99(9):2139-2151.
• [13]Al-Nasiry S, Vercruysse L, Hanssens M, Luyten C, Pijnenborg R: Interstitial trophoblastic cell fusion and E-cadherin immunostaining in the placental bed of normal and hypertensive pregnancies. Placenta 2009, 30(8):719-725.
• [14]Lopez-Novoa JM, Nieto MA: Inflammation and EMT: an alliance towards organ fibrosis and cancer progression. EMBO Mol Med 2009, 1(6-7):303-314.
• [15]Knofler M: Critical growth factors and signalling pathways controlling human trophoblast invasion. Int J Dev Biol 2010, 54(2-3):269-280.
• [16]Zhou BP, Deng J, Xia W, Xu J, Li YM, Gunduz M, Hung MC: Dual regulation of Snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition. Nat Cell Biol 2004, 6(10):931-940.
• [17]Vinas-Castells R, Beltran M, Valls G, Gomez I, Garcia JM, Montserrat-Sentis B, Baulida J, Bonilla F, de Herreros AG, Diaz VM: The hypoxia-controlled FBXL14 ubiquitin ligase targets SNAIL1 for proteasome degradation. J Biol Chem 2010, 285(6):3794-3805.
• [18]Arimoto-Ishida E, Sakata M, Sawada K, Nakayama M, Nishimoto F, Mabuchi S, Takeda T, Yamamoto T, Isobe A, Okamoto Y, et al.: Up-regulation of alpha5-integrin by E-cadherin loss in hypoxia and its key role in the migration of extravillous trophoblast cells during early implantation. Endocrinology 2009, 150(9):4306-4315.
• [19]Butler TM, Elustondo PA, Hannigan GE, MacPhee DJ: Integrin-linked kinase can facilitate syncytialization and hormonal differentiation of the human trophoblast-derived BeWo cell line. Reprod Biol Endocrinol 2009, 7:51. BioMed Central Full Text
• [20]Nakayama H, Scott IC, Cross JC: The transition to endoreduplication in trophoblast giant cells is regulated by the mSNA zinc finger transcription factor. Dev Biol 1998, 199(1):150-163.
• [21]Gultice AD, Selesniemi KL, Brown TL: Hypoxia inhibits differentiation of lineage-specific Rcho-1 trophoblast giant cells. Biol Reprod 2006, 74(6):1041-1050.
• [22]Blechschmidt K, Mylonas I, Mayr D, Schiessl B, Schulze S, Becker KF, Jeschke U: Expression of E-cadherin and its repressor snail in placental tissue of normal, preeclamptic and HELLP pregnancies. Virchows Arch 2007, 450(2):195-202.
• [23]McCarthy FP, Kingdom JC, Kenny LC, Walsh SK: Animal models of preeclampsia; uses and limitations. Placenta 2011, 32(6):413-419.
• [24]Fedorova OV, Kolodkin NI, Agalakova NI, Namikas AR, Bzhelyansky A, St-Louis J, Lakatta EG, Bagrov AY: Antibody to marinobufagenin lowers blood pressure in pregnant rats on a high NaCl intake. J Hypertens 2005, 23(4):835-842.
• [25]Beausejour A, Bibeau K, Lavoie JC, St-Louis J, Brochu M: Placental oxidative stress in a rat model of preeclampsia. Placenta 2007, 28(1):52-58.
• [26]Beausejour A, Auger K, St-Louis J, Brochu M: High-sodium intake prevents pregnancy-induced decrease of blood pressure in the rat. Am J Physiol Heart Circ Physiol 2003, 285(1):H375-H383.
• [27]Vercruysse L, Caluwaerts S, Luyten C, Pijnenborg R: Interstitial trophoblast invasion in the decidua and mesometrial triangle during the last third of pregnancy in the rat. Placenta 2006, 27(1):22-33.
• [28]Lindheimer MD, Taler SJ, Cunningham FG: Hypertension in pregnancy. J Am Soc Hypertens 2008, 2(6):484-494.
• [29]Fedorova OV, Simbirtsev AS, Kolodkin NI, Kotov AY, Agalakova NI, Kashkin VA, Tapilskaya NI, Bzhelyansky A, Reznik VA, Frolova EV, et al.: Monoclonal antibody to an endogenous bufadienolide, marinobufagenin, reverses preeclampsia-induced Na/K-ATPase inhibition and lowers blood pressure in NaCl-sensitive hypertension. J Hypertens 2008, 26(12):2414-2425.
• [30]Fedorova LV, Raju V, El-Okdi N, Shidyak A, Kennedy DJ, Vetteth S, Giovannucci DR, Bagrov AY, Fedorova OV, Shapiro JI, et al.: The cardiotonic steroid hormone marinobufagenin induces renal fibrosis: implication of epithelial-to-mesenchymal transition. Am J Physiol Renal Physiol 2009, 296(4):F922-F934.
• [31]Aplin JD, Jones CJ, Harris LK: Adhesion molecules in human trophoblast--a review. I. Villous trophoblast. Placenta 2009, 30(4):293-298.
• [32]Reuss B, Hellmann P, Dahl E, Traub O, Butterweck A, Grummer R, Winterhager E: Connexins and E-cadherin are differentially expressed during trophoblast invasion and placenta differentiation in the rat. Dev Dyn 1996, 205(2):172-182.
• [33]Vicovac L, Aplin JD: Epithelial-mesenchymal transition during trophoblast differentiation. Acta Anat (Basel) 1996, 156(3):202-216.
• [34]Benian A, Madazli R, Aksu F, Uzun H, Aydin S: Plasma and placental levels of interleukin-10, transforming growth factor-beta1, and epithelial-cadherin in preeclampsia. Obstet Gynecol 2002, 100(2):327-331.
• [35]Li HW, Cheung AN, Tsao SW, Cheung AL, WS O: Expression of e-cadherin and beta-catenin in trophoblastic tissue in normal and pathological pregnancies. Int J Gynecol Pathol 2003, 22(1):63-70.
• [36]Vega S, Morales AV, Ocana OH, Valdes F, Fabregat I, Nieto MA: Snail blocks the cell cycle and confers resistance to cell death. Genes Dev 2004, 18(10):1131-1143.
• [37]Jamora C, Lee P, Kocieniewski P, Azhar M, Hosokawa R, Chai Y, Fuchs E: A signaling pathway involving TGF-beta2 and snail in hair follicle morphogenesis. PLoS Biol 2005, 3(1):e11.
• [38]Valdes F, Alvarez AM, Locascio A, Vega S, Herrera B, Fernandez M, Benito M, Nieto MA, Fabregat I: The epithelial mesenchymal transition confers resistance to the apoptotic effects of transforming growth factor Beta in fetal rat hepatocytes. Mol Cancer Res 2002, 1(1):68-78.
• [39]Georgiades P, Ferguson-Smith AC, Burton GJ: Comparative developmental anatomy of the murine and human definitive placentae. Placenta 2002, 23(1):3-19.
• [40]Nieto MA, Bennett MF, Sargent MG, Wilkinson DG: Cloning and developmental expression of Sna, a murine homologue of the Drosophila snail gene. Development 1992, 116(1):227-237.
• [41]Smith DE, Del Amo Franco F, Gridley T: Isolation of Sna, a mouse gene homologous to the Drosophila genes snail and escargot: its expression pattern suggests multiple roles during postimplantation development. Development 1992, 116(4):1033-1039.
• [42]Zybina TG, Zybina EV: Cell reproduction and genome multiplication in the proliferative and invasive trophoblast cell populations of mammalian placenta. Cell Biol Int 2005, 29(12):1071-1083.
• [43]Zybina TG, Kaufmann P, Frank HG, Freed J, Kadyrov M, Biesterfeld S: Genome multiplication of extravillous trophoblast cells in human placenta in the course of differentiation and invasion into endometrium and myometrium. I. Dynamics of polyploidization. Tsitologiia 2002, 44(11):1058-1067.
• [44]Zybina TG, Frank HG, Biesterfeld S, Kaufmann P: Genome multiplication of extravillous trophoblast cells in human placenta in the course of differentiation and invasion into endometrium and myometrium. II. Mechanisms of polyploidization. Tsitologiia 2004, 46(7):640-648.
• [45]Natale DR, Hemberger M, Hughes M, Cross JC: Activin promotes differentiation of cultured mouse trophoblast stem cells towards a labyrinth cell fate. Dev Biol 2009, 335(1):120-131.
• [46]Nieto MA: The ins and outs of the epithelial to mesenchymal transition in health and disease. Annu Rev Cell Dev Biol 2011, 27:347-376.
• [47]Radisky DC, Levy DD, Littlepage LE, Liu H, Nelson CM, Fata JE, Leake D, Godden EL, Albertson DG, Nieto MA, et al.: Rac1b and reactive oxygen species mediate MMP-3-induced EMT and genomic instability. Nature 2005, 436(7047):123-127.
• [48]Peinado H, Del Carmen Iglesias-de la Cruz M, Olmeda D, Csiszar K, Fong KS, Vega S, Nieto MA, Cano A, Portillo F: A molecular role for lysyl oxidase-like 2 enzyme in snail regulation and tumor progression. EMBO J 2005, 24(19):3446-3458.
• [49]Schietke R, Warnecke C, Wacker I, Schodel J, Mole DR, Campean V, Amann K, Goppelt-Struebe M, Behrens J, Eckardt KU, et al.: The lysyl oxidases LOX and LOXL2 are necessary and sufficient to repress E-cadherin in hypoxia: insights into cellular transformation processes mediated by HIF-1. J Biol Chem 2010, 285(9):6658-6669.
• [50]Postovit LM, Abbott DE, Payne SL, Wheaton WW, Margaryan NV, Sullivan R, Jansen MK, Csiszar K, Hendrix MJ, Kirschmann DA: Hypoxia/reoxygenation: a dynamic regulator of lysyl oxidase-facilitated breast cancer migration. J Cell Biochem 2008, 103(5):1369-1378.
• [51]Hein S, Yamamoto SY, Okazaki K, Jourdan-LeSaux C, Csiszar K, Bryant-Greenwood GD: Lysyl oxidases: expression in the fetal membranes and placenta. Placenta 2001, 22(1):49-57.
• [52]Burton GJ, Hung TH, Jauniaux E: Placental hypoxia, hyperoxia and ischemia-reperfusion injury in preeclampsia. In Preeclampsia Etiology and Clinical Practice. Edited by Belfort LF. Cambridge: Cambridge University Press; 2007:138-151.
• [53]Rajakumar A, Brandon HM, Daftary A, Ness R, Conrad KP: Evidence for the functional activity of hypoxia-inducible transcription factors overexpressed in preeclamptic placentae. Placenta 2004, 25(10):763-769.
• [54]Rajakumar A, Michael HM, Daftary A, Jeyabalan A, Gilmour C, Conrad KP: Proteasomal activity in placentas from women with preeclampsia and intrauterine growth restriction: implications for expression of HIF-alpha proteins. Placenta 2008, 29(3):290-299.
• [55]Huppertz B: Placental pathology in pregnancy complications. Thromb Res 2011, 127(Suppl 3):S96-S99.
• [56]Herse F, Dechend R, Harsem NK, Wallukat G, Janke J, Qadri F, Hering L, Muller DN, Luft FC, Staff AC: Dysregulation of the circulating and tissue-based renin-angiotensin system in preeclampsia. Hypertension 2007, 49(3):604-611.
• [57]Fedorova OV, Tapilskaya NI, Bzhelyansky AM, Frolova EV, Nikitina ER, Reznik VA, Kashkin VA, Bagrov AY: Interaction of Digibind with endogenous cardiotonic steroids from preeclamptic placentae. J Hypertens 2009, 28(2):361-366.
• [58]Williams PJ, Mistry HD, Innes BA, Bulmer JN, Pipkin FB: Expression of AT1R, AT2R and AT4R and their roles in extravillous trophoblast invasion in the human. Placenta 2010, 31(5):448-455.
• [59]Karppanen H, Mervaala E: Sodium intake and hypertension. Prog Cardiovasc Dis 2006, 49(2):59-75.
• [60]Saxena AR, Karumanchi SA, Brown NJ, Royle CM, McElrath TF, Seely EW: Increased sensitivity to angiotensin II is present postpartum in women with a history of hypertensive pregnancy. Hypertension 2010, 55(5):1239-1245.
• [61]St-Louis J, Sicotte B, Beausejour A, Brochu M: Remodeling and angiotensin II responses of the uterine arcuate arteries of pregnant rats are altered by low--and high-sodium intake. Reproduction 2006, 131(2):331-339.
• [62]Beausejour A, Houde V, Bibeau K, Gaudet R, St-Louis J, Brochu M: Renal and cardiac oxidative/nitrosative stress in salt-loaded pregnant rat. Am J Physiol Regul Integr Comp Physiol 2007, 293(4):R1657-R1665.
• [63]Belkacemi L, Nelson DM, Desai M, Ross MG: Maternal undernutrition influences placental-fetal development. Biol Reprod 2010, 83(3):325-331.
• [64]Belkacemi L, Desai M, Michael Nelson D, Ross MG: Altered mitochondrial apoptotic pathway in placentas from undernourished rat gestations. Am J Physiol Regul Integr Comp Physiol 2011, 301(6):R1599-1615.
• [65]Sharp AN, Heazell AE, Crocker IP, Mor G: Placental apoptosis in health and disease. Am J Reprod Immunol 2010, 64(3):159-169.