DOI QR코드

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Zearalenone exposure affects the Wnt/β-catenin signaling pathway and related genes of porcine endometrial epithelial cells in vitro

  • Song, Tingting (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Yang, Weiren (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Huang, Libo (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Yang, Zaibin (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Jiang, Shuzhen (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University)
  • 투고 : 2020.05.03
  • 심사 : 2020.07.27
  • 발행 : 2021.06.01

초록

Objective: Zearalenone (ZEA) has estrogen-like effects. Our previous study has shown that ZEA (0.5 to 1.5 mg/kg) could induce abnormal uterine proliferation through transforming growth factor signaling pathway. To further study the other regulatory networks of uterine hypertrophy caused by ZEA, the potential mechanism of ZEA on porcine endometrial epithelial cells (PECs) was explored by the Illumina Hiseq 2000 sequencing system. Methods: The PECs were treated with ZEA at 0 (ZEA0), 5 (ZEA5), 20 (ZEA20), and 80 (ZEA80) µmol/L for 24 h. The collected cells were subjected to cell cycle, RNA-seq, real-time quantitative polymerase chain reaction, immunofluorescence, and western blot analysis. Results: The proportion of cells in the S and G2 phases decreased (p<0.05), but the proportion of cells in the G1 phase increased (p<0.05) in the ZEA80 treatment. Data analysis revealed that the expression of Wnt pathway-related genes, estrogen-related genes, and mitogen-activated protein kinase pathway-related genes increased (p<0.05), but the expression of genetic stability genes decreased (p<0.05) with increasing ZEA concentrations. The relative mRNA and protein expression of WNT1, β-catenin, glycogen synthase kinase 3β (GSK-3β) were increased (p<0.05) with ZEA increasing, while the relative mRNA and protein expression of cyclin D1 (CCND1) was decreased (p<0.05). Moreover, our immunofluorescence results indicate that β-catenin accumulated around the nucleus from the cell membrane and cytoplasm with increasing ZEA concentrations. Conclusion: In summary, ZEA can activate the Wnt/β-catenin signaling pathway by up-regulating WNT1 and β-catenin expression, to promote the proliferation and development of PECs. At the same time, the up-regulation of GSK-3β and down-regulation of CCND1, as well as the mRNA expression of other pathway related genes indicated that other potential effects of ZEA on the uterine development need further study.

키워드

참고문헌

  1. Othmen ZOB, Golli EE, Abid-Essefi S, Bacha H. Cytotoxicity effects induced by Zearalenone metabolites, α Zearalenol and β Zearalenol, on cultured Vero cells. Toxicology 2008;252:72-7. https://doi.org/10.1016/j.tox.2008.07.065
  2. Rogowska A, Pomastowski P, Rafinska K, et al. A study of zearalenone biosorption and metabolisation by prokaryotic and eukaryotic cells. Toxicon 2019;169:81-90. https://doi.org/10.1016/j.toxicon.2019.09.008
  3. Maragos C. Zearalenone occurrence and human exposure. World Mycotoxin J 2010;3:369-83. https://doi.org/10.3920/WMJ2010.1240
  4. Heneweer M, Houtman R, Poortman J, Groot M, Maliepaard C, Peijnenburg A. Estrogenic effects in the immature rat uterus after dietary exposure to ethinylestradiol and zearalenone using a systems biology approach. Toxicol Sci 2007;99:303-14. https://doi.org/10.1093/toxsci/kfm151
  5. Jiang SZ, Yang ZB, Yang WR, et al. Effects of purified zearalenone on growth performance, organ size, serum metabolites, and oxidative stress in postweaning gilts. J Anim Sci 2011;89:3008-15. https://doi.org/10.2527/jas.2010-3658
  6. Jiang SZ, Yang ZB, Yang WR, et al. Effect of purified zearalenone with or without modified montmorillonite on nutrient availability, genital organs and serum hormones in post-weaning piglets. Livest Sci 2012;144:110-8. https://doi.org/10.1016/j.livsci.2011.11.004
  7. Etienne M, Jemmali M. Effects of zearalenone (F2) on estrous activity and reproduction in gilts. J Anim Sci 1982;55:1-10. https://doi.org/10.2527/jas1982.5511
  8. Chen XX, Yang CW, Huang LB, Niu QS, Jiang SZ, Chi F. Zearalenone altered the serum hormones, morphologic and apoptotic measurements of genital organs in post-weaning gilts. Asian-Australas J Anim Sci 2015;28:171-9. https://doi.org/10.5713/ajas.14.0329
  9. Zhou M, Yang LJ, Yang WR, et al. Effects of zearalenone on the localization and expression of the growth hormone receptor gene in the uteri of post-weaning piglets. Asian-Australas J Anim Sci 2018;31:32-9. https://doi.org/10.5713/ajas.17.0526
  10. Zhou M, Yang L, Chen Y, et al. Comparative study of stress response, growth and development of uteri in post-weaning gilts challenged with zearalenone and estradiol benzoate. J Anim Physiol Anim Nutr 2019;103:1885-94. https://doi.org/10.1111/jpn.13195
  11. Duchartre Y, Kim YM, Kahn M. The Wnt signaling pathway in cancer. Crit Rev Oncol Hematol 2016;99:141-9. https://doi.org/10.1016/j.critrevonc.2015.12.005
  12. Andl T, Reddy ST, Gaddapara T, Millar SE. Wnt signals are required for the initiation of hair follicle development. Dev Cell 2002;2:643-53. https://doi.org/10.1016/S1534-5807(02)00167-3
  13. Lee EY, Choi EJ, Kim JA, et al. Malva verticillata seed extracts upregulate the wnt pathway in human dermal papilla cells. Int J Cosmet Sci 2016;38:148-54. https://doi.org/10.1111/ics.12268
  14. Bai C, Zhang H, Zhang X, Yang W, Li X, Gao Y. MiR-15/16 mediate crosstalk between the MAPK and Wnt/β-catenin pathways during hepatocyte differentiation from amniotic epithelial cells. Biochim Biophys Acta Gene Regul Mech 2019;1862:567-81. https://doi.org/10.1016/j.bbagrm.2019.02.003
  15. Zhang K, Li H, Dong S, et al. Establishment and evaluation of a PRRSV-sensitive porcine endometrial epithelial cell line by transfecting SV40 large T antigen. BMC Vet Res 2019;15:299. https://doi.org/10.1186/s12917-019-2051-1
  16. Yang LJ, Zhou M, Huang LB, et al. Zearalenone-promoted follicle growth through modulation of Wnt-1/β-catenin signaling pathway and expression of estrogen receptor genes in ovaries of postweaning piglets. J Agric Food Chem 2018;66:7899-906. https://doi.org/10.1021/acs.jafc.8b02101
  17. Vlad-Fiegen A, Langerak A, Eberth S, Muller O. The Wnt pathway destabilizes adherens junctions and promotes cell migration via β-catenin and its target gene cyclin D1. FEBS Open Bio 2012;2:26-31. https://doi.org/10.1016/j.fob.2012.02.004
  18. Zeng X, Tamai K, Doble B, et al. A dual-kinase mechanism for Wnt co-receptor phosphorylation and activation. Nature 2005;438:873-7. https://doi.org/10.1038/nature04185
  19. Huang P, Yan R, Zhang X, Wang L, Ke X, Qu Y. Activating Wnt/β-catenin signaling pathway for disease therapy: challenges and opportunities. Pharmacol Ther 2019;196:79-90. https://doi.org/10.1016/j.pharmthera.2018.11.008
  20. Kumawat K, Koopmans T, Gosens R. β-catenin as a regulator and therapeutic target for asthmatic airway remodeling. Expert Opin Ther Targets 2014;18:1023-34. https://doi.org/10.1517/14728222.2014.934813
  21. Luna-Medina R, Cortes-Canteli M, Sanchez-Galiano S, et al. NP031112, a thiadiazolidinone compound, prevents inflammation and neurodegeneration under excitotoxic conditions: potential therapeutic role in brain disorders. J Neurosci 2007;27:5766-76. https://doi.org/10.1523/JNEUROSCI.1004-07.2007
  22. Pazhohan A, Amidi F, Akbari-Asbagh F, et al. The Wnt/β-catenin signaling in endometriosis, the expression of total and active forms of β-catenin, total and inactive forms of glycogen synthase kinase-3β, Wnt7a and Dickkopf-1. Eur J Obstet Gynecol Reprod Biol 2018;220:1-5. https://doi.org/10.1016/j.ejogrb.2017.10.025
  23. Li J, He T, Yuan H. Expression of GSK-3β and E-cadherin in endometrial cancer and its significance. Chongqing Med J 2019;48:288-92.
  24. Koehler A, Schlupf J, Schneider M, Kraft B, Winter C, Kashef J. Loss of Xenopus cadherin-11 leads to increased Wnt/β-catenin signaling and up-regulation of target genes c-myc and cyclin D1 in neural crest. Dev Biol 2013;383:132-45. https://doi.org/10.1016/j.ydbio.2013.08.007
  25. Ye X, Guo Y, Zhang Q, et al. βKlotho suppresses tumor growth in hepatocellular carcinoma by regulating Akt/GSK-3β/cyclin D1 signaling pathway. PLoS One 2013;8:e55615. https://doi.org/10.1371/journal.pone.0055615
  26. Zheng W, Wang B, Li X, et al. Zearalenone promotes cell proliferation or causes cell death? Toxins 2018;10:184. https://doi.org/10.3390/toxins10050184
  27. Kunishige K, Kawate N, Inaba T, Tamada H. Exposure to zearalenone during early pregnancy causes estrogenic multitoxic effects in mice. Reprod Sci 2017;24:421-7. https://doi.org/10.1177/1933719116657194
  28. De Luca A, Maiello MR, D'Alessio A, Pergameno M, Normanno N. The RAS/RAF/MEK/ERK and the PI3K/AKT signalling pathways: role in cancer pathogenesis and implications for therapeutic approaches. Expert Opin Ther Targets 2012;16(Suppl 2):S17-27. https://doi.org/10.1517/14728222.2011.639361
  29. Jiang H, Luo S, Li H. Cdk5 activator-binding protein C53 regulates apoptosis induced by genotoxic stress via modulating the G2/M DNA damage checkpoint. J Biol Chem 2005;280:20651-9. https://doi.org/10.1074/jbc.M413431200
  30. Liu XL, Wu RY, Sun XF, et al. Mycotoxin zearalenone exposure impairs genomic stability of swine follicular granulosa cells in vitro. Int J Biol Sci 2018;14:294-305. https://doi.org/10.7150/ijbs.23898

피인용 문헌

  1. Zearalenone and the Immune Response vol.13, pp.4, 2021, https://doi.org/10.3390/toxins13040248
  2. Impact of Fusarium-Derived Mycoestrogens on Female Reproduction: A Systematic Review vol.13, pp.6, 2021, https://doi.org/10.3390/toxins13060373