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Mitochondrial estrogen receptor β inhibits cell apoptosis via interaction with Bad in a ligand-independent manner

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Abstract

Previous studies reported that estrogen receptor β (ERβ) is localized to mitochondria, whereas little is known about the physiological functions of mitochondrial ERβ. In the present study, we explored the role of mitochondrial ERβ in regulating apoptosis using stable ERβ-expressing and ERβ knockdown cells lines. We found that exogenous ERβ was mainly expressed in mitochondrial but not in nuclear after ERβ overexpression and protected cells from apoptosis induced by hydrogen peroxide (H2O2), ultraviolet (UV), and staurosporine (STS). Moreover, overexpression of ERβ prevented Bax activation, cytochrome c release, caspase-3 activation, and PARP cleavage during apoptosis. Furthermore, knockdown of ERβ significantly suppressed the expression of ERβ in mitochondrial and promoted cell apoptosis induced by H2O2, UV, and STS. Downregulation of ERβ also enhanced Bax activation, cytochrome c release, caspase-3 activation and PARP cleavage. In addition, our study discovered that mitochondrial ERβ interacted with proapoptotic protein Bad in a ligand-independent manner, which suggests that mitochondrial ERβ inhibits Bad, and prevents Bax activation and cytochrome c release. Collectively, the results of this study support that mitochondrial ERβ prevents cell apoptosis via the mitochondrial apoptotic pathway in a ligand-independent manner.

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References

  1. Nadal-Serrano M, Sastre-Serra J, Pons DG, Miro AM, Oliver J, Roca P (2012) The ERalpha/ERbeta ratio determines oxidative stress in breast cancer cell lines in response to 17beta-estradiol. J Cell Biochem 113:3178–3185

    Article  CAS  PubMed  Google Scholar 

  2. Thomas C, Gustafsson JA (2011) The different roles of ER subtypes in cancer biology and therapy. Nat Rev Cancer 11:597–608

    Article  CAS  PubMed  Google Scholar 

  3. Mangelsdorf DJ, Thummel C, Beato M, Herrlich P, Schutz G, Umesono K, Blumberg B, Kastner P, Mark M, Chambon P, Evans RM (1995) The nuclear receptor superfamily: the second decade. Cell 83:835–839

    Article  CAS  PubMed  Google Scholar 

  4. Chang EC, Charn TH, Park SH, Helferich WG, Komm B, Katzenellenbogen JA, Katzenellenbogen BS (2008) Estrogen Receptors alpha and beta as determinants of gene expression: influence of ligand, dose, and chromatin binding. Mol Endocrinol 22:1032–1043

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Le HH, Belcher SM (2010) Rapid signaling actions of environmental estrogens in developing granule cell neurons are mediated by estrogen receptor ss. Endocrinology 151:5689–5699

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Tan XJ, Dai YB, Wu WF, Kim HJ, Barros RP, Richardson TI, Yaden BC, Warner M, McKinzie DL, Krishnan V, Gustafsson JA (2012) Reduction of dendritic spines and elevation of GABAergic signaling in the brains of mice treated with an estrogen receptor beta ligand. Proc Natl Acad Sci USA 109:1708–1712

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Ascenzi P, Bocedi A, Marino M (2006) Structure-function relationship of estrogen receptor alpha and beta: impact on human health. Mol Aspects Med 27:299–402

    Article  CAS  PubMed  Google Scholar 

  8. Majidi M, Al-Wadei HA, Takahashi T, Schuller HM (2007) Nongenomic beta estrogen receptors enhance beta1 adrenergic signaling induced by the nicotine-derived carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in human small airway epithelial cells. Cancer Res 67:6863–6871

    Article  CAS  PubMed  Google Scholar 

  9. Pandini G, Genua M, Frasca F, Squatrito S, Vigneri R, Belfiore A (2007) 17beta-estradiol up-regulates the insulin-like growth factor receptor through a nongenotropic pathway in prostate cancer cells. Cancer Res 67:8932–8941

    Article  CAS  PubMed  Google Scholar 

  10. Cowley SM, Hoare S, Mosselman S, Parker MG (1997) Estrogen receptors alpha and beta form heterodimers on DNA. J Biol Chem 272:19858–19862

    Article  CAS  PubMed  Google Scholar 

  11. Curtis HS, Couse JF, Korach KS (2000) Estrogen receptor transcription and transactivation: estrogen receptor knockout mice: what their phenotypes reveal about mechanisms of estrogen action. Breast Cancer Res 2:345–352

    Article  Google Scholar 

  12. Lubahn DB, Moyer JS, Golding TS, Couse JF, Korach KS, Smithies O (1993) Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor gene. Proc Natl Acad Sci USA 90:11162–11166

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Borras C, Sastre J, Garcia-Sala D, Lloret A, Pallardo FV, Vina J (2003) Mitochondria from females exhibit higher antioxidant gene expression and lower oxidative damage than males. Free Radic Biol Med 34:546–552

    Article  CAS  PubMed  Google Scholar 

  14. Wang X, Simpkins JW, Dykens JA, Cammarata PR (2003) Oxidative damage to human lens epithelial cells in culture: estrogen protection of mitochondrial potential, ATP, and cell viability. Invest Ophthalmol Vis Sci 44:2067–2075

    Article  PubMed  Google Scholar 

  15. Moats RN, Ramirez VD (1998) Rapid uptake and binding of estradiol-17beta-6-(O-carboxymethyl)oxime:125I-labeled BSA by female rat liver. Biol Reprod 58:531–538

    Article  CAS  PubMed  Google Scholar 

  16. Chen JQ, Yager JD, Russo J (2005) Regulation of mitochondrial respiratory chain structure and function by estrogens/estrogen receptors and potential physiological/pathophysiological implications. Biochim Biophys Acta 1746:1–17

    Article  CAS  PubMed  Google Scholar 

  17. Yang SH, Liu R, Perez EJ, Wen Y, Stevens SJ, Valencia T, Brun-Zinkernagel AM, Prokai L, Will Y, Dykens J, Koulen P, Simpkins JW (2004) Mitochondrial localization of estrogen receptor beta. Proc Natl Acad Sci USA 101:4130–4135

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Solakidi S, Psarra AM, Sekeris CE (2005) Differential subcellular distribution of estrogen receptor isoforms: localization of ERalpha in the nucleoli and ERbeta in the mitochondria of human osteosarcoma SaOS-2 and hepatocarcinoma HepG2 cell lines. Biochim Biophys Acta 1745:382–392

    Article  CAS  PubMed  Google Scholar 

  19. Chen JQ, Eshete M, Alworth WL, Yager JD (2004) Binding of MCF-7 cell mitochondrial proteins and recombinant human estrogen receptors alpha and beta to human mitochondrial DNA estrogen response elements. J Cell Biochem 93:358–373

    Article  CAS  PubMed  Google Scholar 

  20. Matsuda K, Nishi M, Takaya H, Kaku N, Kawata M (2008) Intranuclear mobility of estrogen receptor alpha and progesterone receptors in association with nuclear matrix dynamics. J Cell Biochem 103:136–148

    Article  CAS  PubMed  Google Scholar 

  21. Sakamaki J, Daitoku H, Ueno K, Hagiwara A, Yamagata K, Fukamizu A (2011) Arginine methylation of BCL-2 antagonist of cell death (BAD) counteracts its phosphorylation and inactivation by Akt. Proc Natl Acad Sci USA 108:6085–6090

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Huang L, Tang Y, Xing D (2013) Activation of nuclear estrogen receptors induced by low-power laser irradiation via PI3-K/Akt signaling cascade. J Cell Physiol 228:1045–1059

    Article  CAS  PubMed  Google Scholar 

  23. Byer SJ, Eckert JM, Brossier NM, Clodfelder-Miller BJ, Turk AN, Carroll AJ, Kappes JC, Zinn KR, Prasain JK, Carroll SL (2011) Tamoxifen inhibits malignant peripheral nerve sheath tumor growth in an estrogen receptor-independent manner. Neuro Oncol 13:28–41

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Huang L, Wu S, Xing D (2011) High fluence low-power laser irradiation induces apoptosis via inactivation of Akt/GSK3beta signaling pathway. J Cell Physiol 226:588–601

    Article  CAS  PubMed  Google Scholar 

  25. Pedram A, Razandi M, Wallace DC, Levin ER (2006) Functional estrogen receptors in the mitochondria of breast cancer cells. Mol Biol Cell 17:2125–2137

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  26. Yang SH, Sarkar SN, Liu R, Perez EJ, Wang X, Wen Y, Yan LJ, Simpkins JW (2009) Estrogen receptor beta as a mitochondrial vulnerability factor. J Biol Chem 284:9540–9548

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Miller AV, Hicks MA, Nakajima W, Richardson AC, Windle JJ, Harada H (2013) Paclitaxel-induced apoptosis is BAK-dependent, but BAX and BIM-independent in breast tumor. PLoS One 8:e60685

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  28. Zou H, Volonte D, Galbiati F (2012) Interaction of caveolin-1 with Ku70 inhibits Bax-mediated apoptosis. PLoS One 7:e39379

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  29. Andreescu CE, Milojkovic BA, Haasdijk ED, Kramer P, De Jong FH, Krust A, De Zeeuw CI, De Jeu MT (2007) Estradiol improves cerebellar memory formation by activating estrogen receptor beta. J Neurosci 27:10832–10839

    Article  CAS  PubMed  Google Scholar 

  30. Damdimopoulos AE, Spyrou G, Gustafsson JA (2008) Ligands differentially modify the nuclear mobility of estrogen receptors alpha and beta. Endocrinology 149:339–345

    Article  CAS  PubMed  Google Scholar 

  31. Psarra AM, Sekeris CE (2008) Steroid and thyroid hormone receptors in mitochondria. IUBMB Life 60:210–223

    Article  CAS  PubMed  Google Scholar 

  32. Chen JQ, Delannoy M, Cooke C, Yager JD (2004) Mitochondrial localization of ERalpha and ERbeta in human MCF7 cells. Am J Physiol Endocrinol Metab 286:E1011–E1022

    Article  CAS  PubMed  Google Scholar 

  33. Stirone C, Duckles SP, Krause DN, Procaccio V (2005) Estrogen increases mitochondrial efficiency and reduces oxidative stress in cerebral blood vessels. Mol Pharmacol 68:959–965

    Article  CAS  PubMed  Google Scholar 

  34. Robb EL, Stuart JA (2011) Resveratrol interacts with estrogen receptor-beta to inhibit cell replicative growth and enhance stress resistance by upregulating mitochondrial superoxide dismutase. Free Radic Biol Med 50:821–831

    Article  CAS  PubMed  Google Scholar 

  35. Aguirre JI, Plotkin LI, Gortazar AR, Millan MM, O’Brien CA, Manolagas SC, Bellido T (2007) A novel ligand-independent function of the estrogen receptor is essential for osteocyte and osteoblast mechanotransduction. J Biol Chem 282:25501–25508

    Article  CAS  PubMed  Google Scholar 

  36. Xu H, Tai J, Ye H, Kang CB, Yoon HS (2006) The N-terminal domain of tumor suppressor p53 is involved in the molecular interaction with the anti-apoptotic protein Bcl-Xl. Biochem Biophys Res Commun 341:938–944

    Article  CAS  PubMed  Google Scholar 

  37. Chipuk JE, Kuwana T, Bouchier-Hayes L, Droin NM, Newmeyer DD, Schuler M, Green DR (2004) Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science 303:1010–1014

    Article  CAS  PubMed  Google Scholar 

  38. Leu JI, Dumont P, Hafey M, Murphy ME, George DL (2004) Mitochondrial p53 activates Bak and causes disruption of a Bak-Mcl1 complex. Nat Cell Biol 6:443–450

    Article  CAS  PubMed  Google Scholar 

  39. Lin B, Kolluri SK, Lin F, Liu W, Han YH, Cao X, Dawson MI, Reed JC, Zhang XK (2004) Conversion of Bcl-2 from protector to killer by interaction with nuclear orphan receptor Nur77/TR3. Cell 116:527–540

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors thank Dr. Andrew L Wong for excellent technical assistance. They also thank Dr. Steven L. Carroll for kindly providing the ERβ-shRNA construct, and Dr. Ken-Ichi Matsuda and Dr. Akiyoshi Fukamizu for providing the GFP-ERβ and Flag-Bad constructs, respectively.

Conflict of interest

The authors have no financial or personal relationships with other people or organizations that could inappropriately influence their work.

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Authors and Affiliations

  1. Department of Hematology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, China

    Jiayi Liang & Yunxian Chen

  2. Department of Pathology, Medical College of Jinan University, Guangzhou, 510632, China

    Qiang Xie & Xueyun Zhong

  3. Department of Microsurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, China

    Ping Li

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  1. Jiayi Liang
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  2. Qiang Xie
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  3. Ping Li
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  4. Xueyun Zhong
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Corresponding authors

Correspondence to Xueyun Zhong or Yunxian Chen.

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Jiayi Liang and Qiang Xie have contributed equally to this work.

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Liang, J., Xie, Q., Li, P. et al. Mitochondrial estrogen receptor β inhibits cell apoptosis via interaction with Bad in a ligand-independent manner. Mol Cell Biochem 401, 71–86 (2015). https://doi.org/10.1007/s11010-014-2293-y

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  • DOI: https://doi.org/10.1007/s11010-014-2293-y

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