Bisphenol A induces DSB-ATM-p53 signaling leading to cell cycle arrest, senescence, autophagy, stress response, and estrogen release in human fetal lung fibroblasts

Abstract

Experimental and/or epidemiological studies suggest that prenatal exposure to bisphenol A (BPA) may delay fetal lung development and maturation and increase the susceptibility to childhood respiratory disease. However, the underlying mechanisms remain to be elucidated. In our previous study with cultured human fetal lung fibroblasts (HFLF), we demonstrated that 24-h exposure to 1 and 100 µM BPA increased GPR30 protein in the nuclear fraction. Exposure to 100 μM BPA had no effects on cell viability, but increased cytoplasmic expression of ERβ and release of GDF-15, as well as decreased release of IL-6, ET-1, and IP-10 through suppression of NFκB phosphorylation. By performing global gene expression and pathway analysis in this study, we identified molecular pathways, gene networks, and key molecules that were affected by 100, but not 0.01 and 1 µM BPA in HFLF. Using multiple genomic and proteomic tools, we confirmed these changes at both gene and protein levels. Our data suggest that 100 μM BPA increased CYP1B1 and HSD17B14 gene and protein expression and release of endogenous estradiol, which was associated with increased ROS production and DNA double-strand breaks, upregulation of genes and/or proteins in steroid synthesis and metabolism, and activation of Nrf2-regulated stress response pathways. In addition, BPA activated ATM-p53 signaling pathway, resulting in increased cell cycle arrest at G1 phase, senescence and autophagy, and decreased cell proliferation in HFLF. The results suggest that prenatal exposure to BPA at certain concentrations may affect fetal lung development and maturation, and thereby affecting susceptibility to childhood respiratory diseases.

This is a preview of subscription content, access via your institution.

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

References

  1. Abedin MJ, Wang D, McDonnell MA, Lehmann U, Kelekar A (2007) Autophagy delays apoptotic death in breast cancer cells following DNA damage. Cell Death Differ 14:500–510

    CAS  Article  PubMed  Google Scholar 

  2. Adamson IY, Bakowska J, McMillan E, King GM (1990) Accelerated fetal lung maturation by estrogen is associated with an epithelial-fibroblast interaction. In Vitro Cell Dev Biol 26(8):784–790

    CAS  Article  PubMed  Google Scholar 

  3. Alam J, Stewart D, Touchard C, Boinapally S, Choi AM, Cook JL (1999) Nrf2, a Cap’n’Collar transcription factor, regulates induction of the heme oxygenase-1 gene. J Biol Chem 274:26071–26078

    CAS  Article  PubMed  Google Scholar 

  4. Arase S, Ishii K, Igarashi K, Aisaki K, Yoshio Y, Matsushima A, Shimohigashi Y, Arima K, Kanno J, Sugimura Y (2011) Endocrine disrupter bisphenol A increases in situ estrogen production in the mouse urogenital sinus. Biol Reprod 84(4):734–742

    CAS  Article  PubMed  Google Scholar 

  5. Atkinson A, Roy D (1995) In vivo DNA adduct formation by bisphenol A. Environ Mol Mutagen 26(1):60–66

    CAS  Article  PubMed  Google Scholar 

  6. Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B Stat Methodol 57:289–300

    Google Scholar 

  7. Ben-Porath I, Weinberg RA (2005) The signals and pathways activating cellular senescence. Int J Biochem Cell Biol 37(5):961–976

    CAS  Article  PubMed  Google Scholar 

  8. Bièche I, Narjoz C, Asselah T, Vacher S, Marcellin P, Lidereau R, Beaune P, de Waziers I (2007) Reverse transcriptase-PCR quantification of mRNA levels from cytochrome (CYP)1, CYP2 and CYP3 families in 22 different human tissues. Pharmacogenet Genom 17(9):731–742

    Article  Google Scholar 

  9. Bloom MS, Mok-Lin E, Fujimoto VY (2016) Bisphenol A and ovarian steroidogenesis. Fertil Steril 106(4):857–863

    CAS  Article  PubMed  Google Scholar 

  10. Calafat AM, Kuklenyik Z, Reidy JA, Caudill SP, Ekong J, Needham LL (2005) Urinary concentrations of bisphenol A and 4-nonylphenol in a human reference population. Environ Health Perspect 113(4):391–395

    CAS  Article  PubMed  Google Scholar 

  11. Cao XL, Corriveau J, Popovic S, Coughlan MC, Chepelev N, Willmore W, Schrader T, Jin X (2010) Background bisphenol A in experimental materials and its implication to low-dose in vitro study. Chemosphere 81(6):817–820

    CAS  Article  PubMed  Google Scholar 

  12. Cao XL, Zhang J, Goodyer CG, Hayward S, Cooke GM, Curran IH (2012) Bisphenol A in human placental and fetal liver tissues collected from Greater Montreal area (Quebec) during 1998–2008. Chemosphere 89(5):505–511

    CAS  Article  PubMed  Google Scholar 

  13. Cao XL, Perez-Locas C, Robichaud A, Clement G, Popovic S, Dufresne G, Dabeka RW (2015) Levels and temporal trend of bisphenol A in composite food samples from Canadian Total Diet Study 2008–2012. Food Addit Contam Part A Chem Anal. Control Expo Risk Assess 32(12):2154–2160

    CAS  Google Scholar 

  14. Chen Q, Gruber H, Swist E, Pakenham C, Ratnayake WM, Scoggan KA (2009) Influence of dietary phytosterols and phytostanols on diastolic blood pressure and the expression of blood pressure regulatory genes in SHRSP and WKY inbred rats. Br J Nutr 102:93–101

    CAS  Article  PubMed  Google Scholar 

  15. Chen R, Wang B, Chen L, Cai D, Li B, Chen C, Huang E, Liu C, Lin Z, Xie WB, Wang H (2016) DNA damage-inducible transcript 4 (DDIT4) mediates methamphetamine-induced autophagy and apoptosis through mTOR signaling pathway in cardiomyocytes. Toxicol Appl Pharmacol 295:1–11

    CAS  Article  PubMed  Google Scholar 

  16. Cui X, Hwang JT, Qiu J, Blades NJ, Churchill GA (2005) Improved statistical tests for differential gene expression by shrinking variance components estimates. Biostatistics 6:59–75

    Article  PubMed  Google Scholar 

  17. Edlow AG, Chen M, Smith NA, Lu C, McElrath TF (2012) Fetal bisphenol A exposure: concentration of conjugated and unconjugated bisphenol A in amniotic fluid in the second and third trimesters. Reprod Toxicol 34(1):1–7

    CAS  Article  PubMed  Google Scholar 

  18. Ekström L, Johansson M, Rane A (2013) Tissue distribution and relative gene expression of UDP-glucuronosyltransferases (2B7, 2B15, 2B17) in the human fetus. Drug Metab Dispos 41(2):291–295

    Article  PubMed  Google Scholar 

  19. Ganley IG, Lam du H, Wang J, Ding X, Chen S, Jiang X (2009) ULK1.ATG13.FIP200 complex mediates mTOR signaling and is essential for autophagy. J Biol Chem 284(18):12297–12305

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  20. Gao C, Liu L, Ma W, Zhu N, Jiang L, Ren N, Li YF, Kannan K (2016) Bisphenol A in urine of Chinese young adults: concentrations and sources of exposure. Bull Environ Contam Toxicol 96(2):162–167

    CAS  Article  PubMed  Google Scholar 

  21. Hanioka N, Naito T, Narimatsu S (2008) Human UDP-glucuronosyltransferase isoforms involved in bisphenol A glucuronidation. Chemosphere 74(1):33–36

    CAS  Article  PubMed  Google Scholar 

  22. Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains. Exp Cell Res 25:585–621

    CAS  Article  PubMed  Google Scholar 

  23. Hijazi A, Guan H, Cernea M, Yang K (2015) Prenatal exposure to bisphenol A disrupts mouse fetal lung development. FASEB J 29(12):4968–4977

    CAS  Article  PubMed  Google Scholar 

  24. Hines CJ, Jackson MV, Deddens JA, Clark JC, Ye X, Christianson AL, Meadows JW, Calafat AM (2017) Urinary Bisphenol A (BPA) concentrations among workers in industries that manufacture and use BPA in the USA. Ann Work Expo Health 61(2):164–182

    Article  PubMed  PubMed Central  Google Scholar 

  25. Ikezuki Y, Tsutsumi O, Takai Y, Kamei Y, Taketani Y (2002) Determination of bisphenol A concentrations in human biological fluids reveals significant early prenatal exposure. Hum Reprod 17(11):2839–2841

    CAS  Article  PubMed  Google Scholar 

  26. Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP (2003) Summaries of Affymetrix Gene Chip probe level data. Nucleic Acids Res 31(4):e15

    Article  PubMed  PubMed Central  Google Scholar 

  27. Jackson SP (2002) Sensing and repairing DNA double-strand breaks. Carcinogenesis 23(5):687–696

    CAS  Article  PubMed  Google Scholar 

  28. Jaeg JP, Perdu E, Dolo L, Debrauwer L, Cravedi JP, Zalko D (2004) Characterization of new bisphenol a metabolites produced by CD1 mice livermicrosomes and S9 fractions. J Agric Food Chem 52:4935–4942

    CAS  Article  PubMed  Google Scholar 

  29. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19:5720–5728

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  30. Kang JH, Kondo F, Katayama Y (2006) Human exposure to bisphenol A. Toxicology 226(2–3):79–89

    CAS  Article  PubMed  Google Scholar 

  31. Knaak JB, Sullivan LJ (1966) Metabolism of bisphenol a in the rat. Toxicol Appl Pharmacol 8:175–184

    CAS  Article  PubMed  Google Scholar 

  32. Loboda A, Damulewicz M, Pyza E, Jozkowicz A, Dulak J (2016) Role of Nrf2/HO-1 system in development, oxidative stress response and diseases: an evolutionarily conserved mechanism. Cell Mol Life Sci 73(17):3221–3247

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  33. Lucier GW, Sonawane BR, McDaniel OS (1977) Glucuronidation and deglucuronidation reactions in hepatic and extrahepatic tissues during perinatal development. Drug Metab Dispos 5(3):279–287

    CAS  PubMed  Google Scholar 

  34. Mahemuti L, Chen Q, Coughlan MC, Zhang M, Florian M, Mailloux RJ, Cao XL, Scoggan KA, Willmore WG, Jin X (2016) Bisphenol A exposure alters release of immune and developmental modulators and expression of estrogen receptors in human fetal lung fibroblasts. J Environ Sci (China) 48:11–23

    Article  Google Scholar 

  35. Mathiassen SG, De Zio D, Cecconi F (2017) Autophagy and the Cell Cycle: A Complex Landscape. Front Oncol 7:51

    Article  PubMed  PubMed Central  Google Scholar 

  36. Nahar MS, Liao C, Kannan K, Harris C, Dolinoy DC (2015) In utero bisphenol A concentration, metabolism, and global DNA methylation across matched placenta, kidney, and liver in the human fetus. Chemosphere 124:54–60

    CAS  Article  PubMed  Google Scholar 

  37. Nishikawa M, Iwano H, Yanagisawa R, Koike N, Inoue H, Yokota H (2010). Placental transfer of conjugated bisphenol A and subsequent reactivation in the rat fetus. Environ Health Perspect 118(9):1196–1203

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  38. Otterbein LE, Choi AMK (2000). Heme oxygenase: colors of defense against cellular stress. Am J Physiol 279:L1029–1037

    CAS  Google Scholar 

  39. Paine MG, Babu JR, Seibenhener ML (2005) Wooten MW. Evidence for p62 aggregate formation: role in cell survival. FEBS Lett 579:5029–5034

    CAS  Article  PubMed  Google Scholar 

  40. Post M, Barsoumian A, Smith BT (1986) The cellular mechanism of glucocorticoid acceleration of fetal lung maturation. Fibroblast-pneumonocyte factor stimulates choline-phosphate cytidylyltransferase activity. J Biol Chem 261(5):2179–2184

    CAS  PubMed  Google Scholar 

  41. Roy D, Palangat M, Chen C-W, Thomas RD, Colerangle J, Atkinson A, Zhi-Jie Yan Z-J (1997) Biochemical and molecular changes at the cellular level in response to exposure to environmental estrogen-like chemicals. J Toxicol Environ Health 50:1–29

    CAS  Article  PubMed  Google Scholar 

  42. Russo G, Barbato F, Grumetto L (2017) Monitoring of bisphenol A and bisphenol S in thermal paper receipts from the Italian market and estimated transdermal human intake: a pilot study. Sci Total Environ 599–600:68–75

    Article  PubMed  Google Scholar 

  43. Sancar A, Lindsey-Boltz LA, Unsal-Kacmaz K, Linn S (2004) Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Ann Rev Biochem 73:39–85

    CAS  Article  PubMed  Google Scholar 

  44. Seaborn T, Simard M, Provost PR, Piedboeuf B, Tremblay Y (2010) Sex hormone metabolism in lung development and maturation. Trends Endocrinol Metab 21(12):729–738

    CAS  Article  PubMed  Google Scholar 

  45. Spanier AJ, Kahn RS, Kunselman AR, Hornung R, Xu Y, Calafat AM, Lanphear BP (2012) Prenatal exposure to bisphenol A and child wheeze from birth to 3 years of age. Environ Health Perspect 120(6):916–920

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  46. Takahashi O, Oishi S (2000) Disposition of orally administered 2,2-Bis(4-hydroxyphenyl)propane (Bisphenol A) in pregnant rats and the placental transfer to fetuses. Environ Health Perspect 108(10):931–935

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  47. Tonnessen-Murray CA, Lozano G, Jackson JG (2017) The regulation of cellular functions by the p53 protein: cellular senescence. Cold Spring Harb Perspect Med 7(2):a026112

    Article  PubMed  Google Scholar 

  48. Torday JS, Sanchez-Esteban J, Rubin LP (1998) Paracrine mediators of mechanotransduction in lung development. Am J Med Sci 316(3):205–208

    CAS  PubMed  Google Scholar 

  49. Vandenberg LN, Chahoud I, Heindel JJ, Padmanabhan V, Paumgartten FJ, Schoenfelder G (2012) Urinary, circulating, and tissue biomonitoring studies indicate widespread exposure to bisphenol A. Cien Saude Colet 17(2):407–434

    Article  PubMed  Google Scholar 

  50. Williams C, Bondesson M, Krementsov DN, Teuscher C (2014) Gestational bisphenol A exposure and testis development. Endocr Disruptors (Austin) 2(1):e29088

    Article  Google Scholar 

  51. Wu H, Kerr MK, Cui XL, Collin F (2003). MAANOVA: a Software Package for the Analysis of Spotted cDNA Microarray Experiments. Statistics for Biology and Health. Springer, New York, pp 313–341

    Google Scholar 

  52. Yager JD (2000) Endogenous estrogens as carcinogens through metabolic activation. J Natl Cancer Inst Monogr 27:67–73

    CAS  Article  Google Scholar 

  53. Ye X, Zhou X, Needham LL, Calafat AM (2011) In-vitro oxidation of bisphenolA: Is bisphenol A catechol a suitable biomarker for human exposure to bisphenol A? Anal Bioanal Chem 399:1071–1079

    CAS  Article  PubMed  Google Scholar 

  54. Yokota H, Iwano H, Endo M, Kobayashi T, Inoue H, Ikushiro S, Yuasa A (1999) Glucuronidation of the environmental oestrogen bisphenol A by an isoform of UDP-glucuronosyltransferase, UGT2B1, in the rat liver. Biochem J 340(Pt 2):405–409

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  55. Zhang Y, Gaikwad NW, Olson K, Zahid M, Cavalieri EL, Rogan EG (2007) Cytochrome P450 isoforms catalyze formation of catechol estrogen quinones that react with DNA. Metabolism 56(7):887–894

    CAS  Article  PubMed  Google Scholar 

  56. Zhang D, Tang B, Xie X, Xiao YF, Yang SM, Zhang JW (2015) The interplay between DNA repair and autophagy in cancer therapy. Cancer Biol Ther 16(7):1005–1013

    CAS  Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This research was funded by the Chemical Management Plan Research Fund, Government of Canada, Granted to the research team; Discovery Grant (06414-2012) from the Natural Sciences and Engineering Research Council of Canada (NSERC) Granted to WGW and NLC; NSERC-CREATE-REACT (449153-2014) Granted to WGW and LM, Health Canada internal research fund to XJ; and CGS D Scholarship to NLC. The authors thank Ms. Bai Li from University of Ottawa for helping with cell plating and splitting. The authors thank Dr. Ivan Curran in RTRD, Health Canada, for sharing IPA license.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Xiaolei Jin.

Ethics declarations

Conflict of interest

The authors claim no conflict of interests related to the work described in this paper.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Mahemuti, L., Chen, Q., Coughlan, M.C. et al. Bisphenol A induces DSB-ATM-p53 signaling leading to cell cycle arrest, senescence, autophagy, stress response, and estrogen release in human fetal lung fibroblasts. Arch Toxicol 92, 1453–1469 (2018). https://doi.org/10.1007/s00204-017-2150-3

Download citation

Keywords

  • Bisphenol A
  • Human fetal lung fibroblasts
  • ATM signaling
  • DNA double-strand breaks
  • γ-H2AX
  • p53
  • Nrf2
  • ROS
  • Estradiol release
  • CYP1B1
  • HSD17B14
  • Cell cycle arrest
  • G1/S transition
  • Senescence
  • Autophagy
  • Steroid synthesis and metabolism