Apelin and apelin receptor in human placenta: Expression, signalling pathway and regulation of trophoblast JEG‑3 and BeWo cells proliferation and cell cycle

Mlyczyńska,Ewa; Kurowska,Patrycja; Drwal,Eliza; Opydo‑Chanek,Małgorzata; Tworzydło,Wacław; Kotula‑Balak,Małgorzata; Rak,Agnieszka

doi:10.3892/ijmm.2020.4452

Apelin and apelin receptor in human placenta: Expression, signalling pathway and regulation of trophoblast JEG‑3 and BeWo cells proliferation and cell cycle

Authors:
- Ewa Mlyczyńska
- Patrycja Kurowska
- Eliza Drwal
- Małgorzata Opydo‑Chanek
- Wacław Tworzydło
- Małgorzata Kotula‑Balak
- Agnieszka Rak
View Affiliations

Affiliations: Department of Physiology and Toxicology of Reproduction, Institute of Zoology and Biomedical Research, Jagiellonian University in Krakow, 30‑387 Krakow, Poland, Department of Experimental Haematology, Institute of Zoology and Biomedical Research, Jagiellonian University in Krakow, 30‑387 Krakow, Poland, Department of Developmental Biology and Invertebrate Morphology, Institute of Zoology and Biomedical Research, Jagiellonian University in Krakow, 30‑387 Krakow, Poland, University Centre of Veterinary Medicine UJ‑UR, University of Agriculture in Krakow, 30‑059 Krakow, Poland
Published online on: January 7, 2020 https://doi.org/10.3892/ijmm.2020.4452
Pages: 691-702
Copyright: © Mlyczyńska et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Placentation requires the production of numerous growth factors, hormones and transcription factors. Many of them, like the adipose tissue‑derived leptin or adiponectin, have been identified in the placenta and their role has been established in the proliferation and subsequent development of the placenta. Apelin is another adipokine known for proliferative effects in different cell types. PCR, immunoblotting and immunocytochemistry were used to study mRNA and protein expression of apelin and its receptor (APJ) in syncytiotrophoblast (BeWo) and cytotrophoblast (JEG‑3) cells as well in immunohistochemistry in human normal placenta slides. The effect of apelin on cell proliferation study was investigated by alamarBlue® and Cell Counting Kit‑8 assays, the cell cycle by the flow cytometry method and the protein expression of cyclins and phosphorylation level of extracellular signal‑regulated kinases (ERK)1/2, phosphatidylinositol 3'‑kinase/protein kinase B (Akt), signal transducer and activator of transcription 3 (Stat3) and 5'‑monophosphate‑activated protein kinase (AMPKα) were studied by western blotting. Apelin was increased in JEG‑3 compared with in BeWo cells, while APJ was the same in both placenta cell lines. Immunocytochemical analyses revealed high cytoplasmic and/or membrane apelin localisation in JEG‑3, while BeWo cells exhibited markedly weaker apelin signal in the cytoplasm. Apelin increased cell proliferation as well as the percentage of cells in the G2/M phase of the cell cycle, cyclin proteins and the expression of all kinases mentioned above. In conclusion, apelin by promotion of trophoblast cell proliferation by APJ and ERK1/2, Stat3 and AMPKα signalling could be a new important adipokine in the regulation of early placental development.

View Figures

View References

1	Maltepe E, Bakardjiev AI and Fisher SJ: The placenta: Transcriptional, epigenetic, and physiological integration during development. J Clin Invest. 120:1016–1025. 2010. View Article : Google Scholar : PubMed/NCBI
2	Scifres CM and Nelson DM: Intrauterine growth restriction, human placental development and trophoblast cell death. J Physiol. 587:3453–3458. 2009. View Article : Google Scholar : PubMed/NCBI
3	Cobellis L, De Falco M, Mastrogiacomo A, Giraldi D, Dattilo D, Scaffa C, Colacurci N and De Luca A: Modulation of apelin and APJ receptor in normal and preeclampsia-complicated placentas. Histol Histopathol. 22:1–8. 2007.
4	Reus AD, El-Harbachi H, Rousian M, Willemsen SP, Steegers-Theunissen RP, Steegers EA and Exalto N: Early first-trimester trophoblast volume in pregnancies that result in live birth or miscarriage. Ultrasound Obstet Gynecol. 42:577–584. 2013. View Article : Google Scholar : PubMed/NCBI
5	Murphy VE, Smith R, Giles WB and Clifton VL: Endocrine regulation of human fetal growth: The role of the mother, placenta, and fetus. Endocr Rev. 27:141–169. 2006. View Article : Google Scholar : PubMed/NCBI
6	Briana DD and Malamitsi-Puchner A: Intrauterine growth restriction and adult disease: The role of adipocytokines. Eur J Endocrinol. 160:337–347. 2009. View Article : Google Scholar
7	Knöfler M and Pollheimer J: IFPA Award in Placentology lecture: Molecular regulation of human trophoblast invasion. Placenta. 33(Suppl): S55–S62. 2012. View Article : Google Scholar :
8	Forbes K, Westwood M, Baker PN and Aplin JD: Insulin-like growth factor I and II regulate the life cycle of trophoblast in the developing human placenta. Am J Physiol Cell Physiol. 294:C1313–C1322. 2008. View Article : Google Scholar : PubMed/NCBI
9	Caüzac M, Czuba D, Girard J and Hauguel-de Mouzon S: Transduction of leptin growth signals in placental cells is independent of JAK-STAT activation. Placenta. 24:378–384. 2003. View Article : Google Scholar : PubMed/NCBI
10	Schulz LC and Widmaier EP: The effect of leptin on mouse trophoblast cell invasion. Biol Reprod. 71:1963–1967. 2004. View Article : Google Scholar : PubMed/NCBI
11	Magariños MP, Sánchez-Margalet V, Kotler M, Calvo JC and Varone CL: Leptin promotes cell proliferation and survival of trophoblastic cells. Biol Reprod. 76:203–210. 2007. View Article : Google Scholar
12	Benaitreau D, Dieudonné MN, Dos Santos E, Leneveu MC, Mazancourt Pd and Pecquery R: Antiproliferative effects of adiponectin on human trophoblastic cell lines JEG-3 and BeWo. Biol Reprod. 80:1107–1114. 2009. View Article : Google Scholar : PubMed/NCBI
13	De Falco M, De Luca L, Onori N, Cavallotti I, Artigiano F, Esposito V, De Luca B, Laforgia V, Groeger AM and De Luca A: Apelin expression in normal human tissues. In Vivo. 16:333–336. 2002.PubMed/NCBI
14	Kurowska P, Barbe A, Różycka M, Chmielińska J, Dupont J and Rak A: Apelin in reproductive physiology and pathology of different species: A critical review. Int J Endocrinol. 2018:91704802018. View Article : Google Scholar : PubMed/NCBI
15	Boucher J, Masri B, Daviaud D, Gesta S, Guigné C, Mazzucotelli A, Castan-Laurell I, Tack I, Knibiehler B, Carpéné C, et al: Apelin, a newly identified adipokine up-regulated by insulin and obesity. Endocrinology. 146:1764–1771. 2005. View Article : Google Scholar : PubMed/NCBI
16	Lee DK, Cheng R, Nguyen T, Fan T, Kariyawasam AP, Liu Y, Osmond DH, George SR and O'Dowd BF: Characterization of apelin, the ligand for the APJ receptor. J Neurochem. 74:34–41. 2000. View Article : Google Scholar : PubMed/NCBI
17	Bertrand C, Valet P and Castan-Laurell I: Apelin and energy metabolism. Front Physiol. 6:1152015. View Article : Google Scholar : PubMed/NCBI
18	Tatemoto K, Takayama K, Zou MX, Kumaki I, Zhang W, Kumano K and Fujimiya M: The novel peptide apelin lowers blood pressure via a nitric oxide-dependent mechanism. Regul Pept. 99:87–92. 2001. View Article : Google Scholar : PubMed/NCBI
19	Kidoya H and Takakura N: Biology of the apelin-APJ axis in vascular formation. J Biochem. 152:125–131. 2012. View Article : Google Scholar : PubMed/NCBI
20	Tatemoto K, Hosoya M, Habata Y, Fujii R, Kakegawa T, Zou MX, Kawamata Y, Fukusumi S, Hinuma S, Kitada C, et al: Isolation and characterization of a novel endogenous peptide ligand for the human APJ receptor. Biochem Biophys Res Commun. 251:471–476. 1998. View Article : Google Scholar : PubMed/NCBI
21	Yamaleyeva LM, Chappell MC, Brosnihan KB, Anton L, Caudell DL, Shi S, McGee C, Pirro N, Gallagher PE, Taylor RN, et al: Downregulation of apelin in the human placental chorionic villi from preeclamptic pregnancies. Am J Physiol Endocrinol Metab. 309:E852–E860. 2015. View Article : Google Scholar : PubMed/NCBI
22	Charo DN, Ho M, Fajardo G, Kawana M, Kundu RK, Sheikh AY, Finsterbach TP, Leeper NJ, Ernst KV, Chen MM, et al: Endogenous regulation of cardiovascular function by apelin-APJ. Am J Physiol Heart Circ Physiol. 297:H1904–H1913. 2009. View Article : Google Scholar : PubMed/NCBI
23	Van Mieghem T, van Bree R, Van Herck E, Pijnenbor R, Deprest J and Verhaeghe J: Maternal apelin physiology during rat pregnancy: The role of the placenta. Placenta. 31:725–730. 2010. View Article : Google Scholar : PubMed/NCBI
24	Hanssens A, Marx-Deseure S, Lecoutre L, Butruille A, Fournel C, Knauf C, Besengez C, Breton L, Storme P, Deruelle, et al: Maternal obesity alters the apelinergic system at the feto-maternal interface. Placenta. 39:41–44. 2016. View Article : Google Scholar : PubMed/NCBI
25	Malamitsi-Puchner A, Gourgiotis D, Boutsikou M, Baka S, Hassiakos D and Briana DD: Circulating apelin concentrations in mother/infant pairs at term. Acta Paediatr. 96:1751–1754. 2007. View Article : Google Scholar : PubMed/NCBI
26	Van Mieghem T, Doherty A, Baczyk D, Drewlo S, Baud D, Carvalho J and Kingdom J: Apelin in normal pregnancy and pregnancies complicated by placental insufficiency. Reprod Sci. 23:1037–1043. 2016. View Article : Google Scholar : PubMed/NCBI
27	Cheng JQ, Lindsley CW, Cheng GZ, Yang H and Nicosia SV: The Akt/PKB pathway: Molecular target for cancer drug discovery. Oncogene. 24:7482–7492. 2005. View Article : Google Scholar : PubMed/NCBI
28	Thompson N and Lyons J: Recent progress in targeting the Raf/MEK/ERK pathway with inhibitors in cancer drug discovery. Curr Opin Pharmacol. 5:350–356. 2005. View Article : Google Scholar : PubMed/NCBI
29	Pereira de Sousa FL, Chaiwangyen W, Morales-Prieto DM, Ospina-Prieto S, Weber M, Photini SM, Sass N, Daher S, Schleussner E and Markert UR: Involvement of STAT1 in proliferation and invasiveness of trophoblastic cells. Reprod Biol. 17:218–224. 2017. View Article : Google Scholar : PubMed/NCBI
30	Qin D, Zheng XX and Jiang YR: Apelin-13 induces proliferation, migration, and collagen I mRNA expression in human RPE cells via PI3K/Akt and MEK/Erk signaling pathways. Mol Vis. 19:2227–2236. 2013.PubMed/NCBI
31	Li Y, Bai YJ, Jiang YR, Yu WZ, Shi X, Chen L, Feng J and Sun GB: Apelin-13 is an early promoter of cytoskeleton and tight junction in diabetic macular edema via PI-3K/Akt and MAPK/Erk and MAPK/Erk signaling pathways. Biomed Res Int. 2018:32425742018.
32	Różycka M, Kurowska P, Grzesiak M, Kotula-Balak M, Tworzydło W, Rame C, Gregoraszczuk E, Dupont J and Rak A: Apelin and apelin receptor at different stages of corpus luteum development and effect of apelin on progesterone secretion and 3β-hydroxysteroid dehydrogenase (3β-HSD) in pigs. Anim Reprod Sci. 92:251–260. 2018. View Article : Google Scholar
33	Rak A, Drwal E, Rame C, Knapczyk-Stwora K, Słomczyńska M, Dupont J and Gregoraszczuk EL: Expression of apelin and apelin receptor (APJ) in porcine ovarian follicles and in vitro effect of apelin on steroidogenesis and proliferation through APJ activation and different signaling pathway. Theriogenology. 96:126–135. 2017. View Article : Google Scholar : PubMed/NCBI
34	Jasaszwili M, Wojciechowicz T, Billert M, Strowski MZ, Nowak KW and Skrzypski M: Effects of adropin on proliferation and differentiation of 3T3-L1 cells and rat primary preadipocytes. Mol Cell Endocrinol. 496:1105322019. View Article : Google Scholar : PubMed/NCBI
35	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
36	Rebut-Bonneton C, Segond N, Demignon J, Porquet D and Evain-Brion D: Effects of calcitonin on human trophoblastic cells in culture: absence of autocrine control. Mol Cell Endocrinol. 85:65–71. 1992. View Article : Google Scholar : PubMed/NCBI
37	Zygmunt M, Hahn D, Munstedt K, Bischof P and Lang U: Invasion of cytotrophoblastic JEG-3 cells is stimulated by hCG in vitro. Placenta. 19:587–593. 1998. View Article : Google Scholar : PubMed/NCBI
38	Standley PR and Standley CA: Identification of a functional Na+/Mg2+ exchanger in human trophoblast cells. Am J Hypertens. 15:565–570. 2002. View Article : Google Scholar : PubMed/NCBI
39	Wysocka MB, Pietraszek-Gremplewicz K and Nowak D: The role of apelin in cardiovascular diseases, obesity and cancer. Front Physiol. 9:5572018. View Article : Google Scholar : PubMed/NCBI
40	Wang G, Anini Y, Wei W, Qi X, OCarroll AM, Mochizuki T, Wang HQ, Hellmich MR, Englander EW and Greeley GH Jr: Apelin, a new enteric peptide, localization in the gastrointestinal tract, ontogeny, stimulation of gastric cell proliferation and of cholecystokinin secretion. Endocrinology. 145:1342–1348. 2004. View Article : Google Scholar
41	Masri B, Knibiehler B and Audigier Y: Apelin signaling: A promising pathway from cloning to pharmacology. Cell Signal. 17:415–426. 2005. View Article : Google Scholar
42	Kourtis A, Gkiomisi A, Mouzaki M, Makedou K, Anastasilakis AD, Toulis KA, Gerou S, Gavana E and Agorastos T: Apelin levels in normal pregnancy. Clin Endocrinol (Oxf). 75:367–371. 2011. View Article : Google Scholar
43	Yin L, Zhang P, Li C, Si J, Wang Y, Zhang X, Zhang D, Zhang H and Lin C: Apelin-13 promotes cell proliferation in the H9c2 cardiomyoblast cell line by triggering extracellular signal-regulated kinase 1/2 and protein kinase B phosphorylation. Mol Med Rep. 17:447–451. 2018.
44	Li F, Li L, Qin X, Pan W, Feng F, Chen F, Zhu B, Liao D, Tanowitz H, Albanese C and Chen L: Apelin-induced vascular smooth muscle cell proliferation: The regulation of cyclin D1. Front Biosci. 13:3786–3792. 2008. View Article : Google Scholar : PubMed/NCBI
45	Sorli SC, van den Berghe L, Masri B, Knibiehler B and Audigier Y: Therapeutic potential of interfering with apelin signalling. Drug Discov Today. 11:1100–1106. 2006. View Article : Google Scholar : PubMed/NCBI
46	Sánchez I and Dynlacht BD: New insights into cyclins, CDKs, and cell cycle control. Semin Cell Dev Biol. 16:311–321. 2005. View Article : Google Scholar : PubMed/NCBI
47	Bertoli C, Skotheim JM and de Bruin RA: Control of cell cycle transcription during G1 and S phases. Nat Rev Mol Cell Biol. 14:518–528. 2013. View Article : Google Scholar : PubMed/NCBI
48	Foster DA, Yellen P, Xu L and Saqcena M: Regulation of G1 cell cycle progression: Distinguishing the restriction point from a nutrient-sensing cell growth checkpoint(s). Genes Cancer. 11:1124–1131. 2010. View Article : Google Scholar
49	Shuang L, Jidong W, Hongjuan P and Zhenwei Y: Effects of apelin on proliferation and apoptosis in rat ovarian granulosa cells. Clin Exp Obstet Gynecol. 43:409–413. 2016.PubMed/NCBI
50	Zhang J, Liu Q, Hu X, Fang Z, Huang F, Tang L and Zhou S: Apelin/APJ signaling promotes hypoxia-induced proliferation of endothelial progenitor cells via phosphoinositide-3 kinase/Akt signaling. Mol Med Rep. 12:3829–3834. 2015. View Article : Google Scholar : PubMed/NCBI
51	Peng X, Li F, Wang P, Jia S, Sun L and Huo H: Apelin-13 induces MCF-7 cell proliferation and invasion via phosphorylation of ERK1/2. Int J Mol Med. 36:733–738. 2015. View Article : Google Scholar : PubMed/NCBI
52	Yang L, Su T, Lv D, Xie F, Liu W, Cao J, Sheikh IA, Qin X, Li L and Chen L: ERK1/2 mediates lung adenocarcinoma cell proliferation and autophagy induced by apelin-13. Acta Biochim Biophys Sin (Shanghai). 6:100–111. 2014. View Article : Google Scholar