Dual Epigenetic Regulation of ERα36 Expression in Breast Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. Screening of microRNAs that Target ERα36 but not ERα66
2.2. hsa-miR-136-5p Under/over Expression Modulates ERα36 mRNA Expression Level
2.3. The Demethylating Agent DAC Stimulates hsa-miR-136-5p and Represses ERα36 Expression
2.4. DAC Counteracts OHT-dependent Stimulation of ERα36 Expression
2.5. ERα36 Expression May be Modulated by the Methylation Status of Its Promoter, In Vivo and In Vitro
2.6. DAC Directly Targets ERα36 Genomic Sequence
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Plasmids
4.3. Cell Culture
4.4. Transfection
4.5. Luciferase/Alkaline Phosphatase Assays
4.6. MicroRNA Extraction and Reverse Transcription
4.7. Long RNAs RT and Real-time PCR Analysis
4.8. Western Immunoblotting
4.9. Patients and Samples
4.10. Bisulfite Sequencing
4.11. DNA Methylation Analysis or ERα36 Promoter Sequence
4.12. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Szostakowska, M.; Trębińska-Stryjewska, A.; Grzybowska, E.A.; Fabisiewicz, A. Resistance to endocrine therapy in breast cancer: Molecular mechanisms and future goals. Breast Cancer Res. Treat. 2019, 173, 489–497. [Google Scholar] [CrossRef]
- Wang, Q.; Jiang, J.; Ying, G.; Xie, X.-Q.; Zhang, X.; Xu, W.; Zhang, X.; Song, E.; Bu, H.; Ping, Y.-F.; et al. Tamoxifen enhances stemness and promotes metastasis of ERα36+ breast cancer by upregulating ALDH1A1 in cancer cells. Cell Res. 2018, 28, 336–358. [Google Scholar] [CrossRef]
- Wang, Z.; Zhang, X.; Shen, P.; Loggie, B.W.; Chang, Y.; Deuel, T.F. A variant of estrogen receptor-α, hER-α36: Transduction of estrogen- and antiestrogen-dependent membrane-initiated mitogenic signaling. Proc. Natl. Acad. Sci. USA 2006, 103, 9063–9068. [Google Scholar] [CrossRef]
- Zhang, X.; Wang, Z.-Y. Estrogen receptor-α variant, ER-α36, is involved in tamoxifen resistance and estrogen hypersensitivity. Endocrinology 2013, 154, 1990–1998. [Google Scholar] [CrossRef]
- Li, G.; Zhang, J.; Jin, K.; He, K.; Zheng, Y.; Xu, X.; Wang, H.; Wang, H.; Li, Z.; Yu, X.; et al. Estrogen receptor-α36 is involved in development of acquired tamoxifen resistance via regulating the growth status switch in breast cancer cells. Mol. Oncol. 2013, 7, 611–624. [Google Scholar] [CrossRef] [PubMed]
- Wang, Z.; Zhang, X.; Shen, P.; Loggie, B.W.; Chang, Y.; Deuel, T.F. Identification, cloning, and expression of human estrogen receptor-alpha36, a novel variant of human estrogen receptor-alpha66. Biochem. Biophys. Res. Commun. 2005, 336, 1023–1027. [Google Scholar] [CrossRef]
- Chaudhri, R.A.; Hadadi, A.; Lobachev, K.S.; Schwartz, Z.; Boyan, B.D. Estrogen receptor-alpha 36 mediates the anti-apoptotic effect of estradiol in triple negative breast cancer cells via a membrane-associated mechanism. Biochim. Biophys. Acta 2014, 1843, 2796–2806. [Google Scholar] [CrossRef] [Green Version]
- Omarjee, S.; Jacquemetton, J.; Poulard, C.; Rochel, N.; Dejaegere, A.; Chebaro, Y.; Treilleux, I.; Marangoni, E.; Corbo, L.; Romancer, M.L. The molecular mechanisms underlying the ERα-36-mediated signaling in breast cancer. Oncogene 2017, 36, 2503–2514. [Google Scholar] [CrossRef]
- Chaudhri, R.A.; Schwartz, N.; Elbaradie, K.; Schwartz, Z.; Boyan, B.D. Role of ERα36 in membrane-associated signaling by estrogen. Steroids 2014, 81, 74–80. [Google Scholar] [CrossRef] [PubMed]
- Thiebaut, C.; Chamard-Jovenin, C.; Chesnel, A.; Morel, C.; Djermoune, E.-H.; Boukhobza, T.; Dumond, H. Mammary epithelial cell phenotype disruption in vitro and in vivo through ERalpha36 overexpression. PLoS ONE 2017, 12, e0173931. [Google Scholar] [CrossRef] [PubMed]
- Wang, Z.-Y.; Yin, L. Estrogen receptor alpha-36 (ER-α36): A new player in human breast cancer. Mol. Cell. Endocrinol. 2015, 418, 193–206. [Google Scholar] [CrossRef]
- Sołtysik, K.; Czekaj, P. ERα36--Another piece of the estrogen puzzle. Eur. J. Cell Biol. 2015, 94, 611–625. [Google Scholar] [CrossRef]
- Chamard-Jovenin, C.; Jung, A.C.; Chesnel, A.; Abecassis, J.; Flament, S.; Ledrappier, S.; Macabre, C.; Boukhobza, T.; Dumond, H. From ERα66 to ERα36: A generic method for validating a prognosis marker of breast tumor progression. BMC Syst. Biol. 2015, 9, 28. [Google Scholar] [CrossRef]
- Zou, Y.; Ding, L.; Coleman, M.; Wang, Z. Estrogen receptor-alpha (ER-α) suppresses expression of its variant ER-α36. FEBS Lett. 2009, 583, 1368–1374. [Google Scholar] [CrossRef]
- Yin, L.; Wang, Z.-Y. Roles of the ER-α36-EGFR/HER2 positive regulatory loops in tamoxifen resistance. Steroids 2016, 111, 95–99. [Google Scholar] [CrossRef]
- Chamard-Jovenin, C.; Thiebaut, C.; Chesnel, A.; Bresso, E.; Morel, C.; Smail-Tabbone, M.; Devignes, M.-D.; Boukhobza, T.; Dumond, H. Low-Dose Alkylphenol Exposure Promotes Mammary Epithelium Alterations and Transgenerational Developmental Defects, But Does Not Enhance Tumorigenic Behavior of Breast Cancer Cells. Front. Endocrinol. 2017, 8, 272. [Google Scholar] [CrossRef]
- Piletič, K.; Kunej, T. MicroRNA epigenetic signatures in human disease. Arch. Toxicol. 2016, 90, 2405–2419. [Google Scholar] [CrossRef]
- Yan, M.; Li, X.; Tong, D.; Han, C.; Zhao, R.; He, Y.; Jin, X. miR-136 suppresses tumor invasion and metastasis by targeting RASAL2 in triple-negative breast cancer. Oncol. Rep. 2016, 36, 65–71. [Google Scholar] [CrossRef] [Green Version]
- Huan, J.; Xing, L.; Lin, Q.; Xui, H.; Qin, X. Long noncoding RNA CRNDE activates Wnt/β-catenin signaling pathway through acting as a molecular sponge of microRNA-136 in human breast cancer. Am. J. Transl. Res. 2017, 9, 1977–1989. [Google Scholar]
- Guo, T.; Pan, G. MicroRNA-136 functions as a tumor suppressor in osteosarcoma via regulating metadherin. Cancer Biomark. Sect. Dis. Markers 2018, 22, 79–87. [Google Scholar] [CrossRef]
- Deng, G.; Gao, Y.; Cen, Z.; He, J.; Cao, B.; Zeng, G.; Zong, S. miR-136-5p Regulates the Inflammatory Response by Targeting the IKKβ/NF-κB/A20 Pathway After Spinal Cord Injury. Cell. Physiol. Biochem. Int. J. Exp. Cell. Physiol. Biochem. Pharmacol. 2018, 50, 512–524. [Google Scholar] [CrossRef]
- Xie, Z.-C.; Li, T.-T.; Gan, B.-L.; Gao, X.; Gao, L.; Chen, G.; Hu, X.-H. Investigation of miR-136-5p key target genes and pathways in lung squamous cell cancer based on TCGA database and bioinformatics analysis. Pathol. Res. Pract. 2018, 214, 644–654. [Google Scholar] [CrossRef]
- Chen, P.; Zhao, L.; Pan, X.; Jin, L.; Lin, C.; Xu, W.; Xu, J.; Guan, X.; Wu, X.; Wang, Y.; et al. Tumor suppressor microRNA-136-5p regulates the cellular function of renal cell carcinoma. Oncol. Lett. 2018, 15, 5995–6002. [Google Scholar] [CrossRef]
- Li, T.-T.; Gao, X.; Gao, L.; Gan, B.-L.; Xie, Z.-C.; Zeng, J.-J.; Chen, G. Role of upregulated miR-136-5p in lung adenocarcinoma: A study of 1242 samples utilizing bioinformatics analysis. Pathol. Res. Pract. 2018, 214, 750–766. [Google Scholar] [CrossRef]
- Kozomara, A.; Griffiths-Jones, S. miRBase: Annotating high confidence microRNAs using deep sequencing data. Nucleic Acids Res. 2014, 42, D68–D73. [Google Scholar] [CrossRef]
- Kozomara, A.; Griffiths-Jones, S. miRBase: Integrating microRNA annotation and deep-sequencing data. Nucleic Acids Res. 2011, 39, D152–D157. [Google Scholar] [CrossRef]
- Griffiths-Jones, S.; Grocock, R.J.; van Dongen, S.; Bateman, A.; Enright, A.J. miRBase: microRNA sequences, targets and gene nomenclature. Nucleic Acids Res. 2006, 34, D140–D144. [Google Scholar] [CrossRef]
- Griffiths-Jones, S.; Saini, H.K.; van Dongen, S.; Enright, A.J. miRBase: Tools for microRNA genomics. Nucleic Acids Res. 2008, 36, D154–D158. [Google Scholar] [CrossRef]
- Panwar, B.; Omenn, G.S.; Guan, Y. miRmine: A database of human miRNA expression profiles. Bioinform. Oxf. Engl. 2017, 33, 1554–1560. [Google Scholar] [CrossRef]
- Gulyaeva, L.F.; Kushlinskiy, N.E. Regulatory mechanisms of microRNA expression. J. Transl. Med. 2016, 14, 143. [Google Scholar] [CrossRef]
- Eggermann, T.; Perez de Nanclares, G.; Maher, E.R.; Temple, I.K.; Tümer, Z.; Monk, D.; Mackay, D.J.G.; Grønskov, K.; Riccio, A.; Linglart, A.; et al. Imprinting disorders: A group of congenital disorders with overlapping patterns of molecular changes affecting imprinted loci. Clin. Epigenetics 2015, 7, 123. [Google Scholar] [CrossRef]
- Uppal, A.; Wightman, S.C.; Mallon, S.; Oshima, G.; Pitroda, S.P.; Zhang, Q.; Huang, X.; Darga, T.E.; Huang, L.; Andrade, J.; et al. 14q32-encoded microRNAs mediate an oligometastatic phenotype. Oncotarget 2015, 6, 3540–3552. [Google Scholar] [CrossRef]
- Xu, Z.; Huang, B.; Liu, J.; Wu, X.; Luo, N.; Wang, X.; Zheng, X.; Pan, X. Combinatorial anti-proliferative effects of tamoxifen and naringenin: The role of four estrogen receptor subtypes. Toxicology 2018, 410, 231–246. [Google Scholar] [CrossRef]
- Ajj, H.; Chesnel, A.; Pinel, S.; Plénat, F.; Flament, S.; Dumond, H. An alkylphenol mix promotes seminoma derived cell proliferation through an ERalpha36-mediated mechanism. PLoS ONE 2013, 8, e61758. [Google Scholar] [CrossRef]
- Ariazi, E.A.; Taylor, J.C.; Black, M.A.; Nicolas, E.; Slifker, M.J.; Azzam, D.J.; Boyd, J. A New Role for ERα: Silencing via DNA Methylation of Basal, Stem Cell, and EMT Genes. Mol. Cancer Res. 2017, 15, 152–164. [Google Scholar] [CrossRef]
- Chaudhri, R.A.; Olivares-Navarrete, R.; Cuenca, N.; Hadadi, A.; Boyan, B.D.; Schwartz, Z. Membrane estrogen signaling enhances tumorigenesis and metastatic potential of breast cancer cells via estrogen receptor-α36 (ERα36). J. Biol. Chem. 2012, 287, 7169–7181. [Google Scholar] [CrossRef]
- Shi, L.; Dong, B.; Li, Z.; Lu, Y.; Ouyang, T.; Li, J.; Wang, T.; Fan, Z.; Fan, T.; Lin, B.; et al. Expression of ER-{alpha}36, a novel variant of estrogen receptor {alpha}, and resistance to tamoxifen treatment in breast cancer. J. Clin. Oncol. 2009, 27, 3423–3429. [Google Scholar] [CrossRef]
- Fleischer, T.; Tekpli, X.; Mathelier, A.; Wang, S.; Nebdal, D.; Dhakal, H.P.; Sahlberg, K.K.; Schlichting, E.; Børresen-Dale, A.-L.; Borgen, E.; et al. DNA methylation at enhancers identifies distinct breast cancer lineages. Nat. Commun. 2017, 8, 1379. [Google Scholar] [CrossRef]
- Xue, X.; Yang, Y.A.; Zhang, A.; Fong, K.; Kim, J.; Song, B.; Li, S.; Zhao, J.C.; Yu, J. LncRNA HOTAIR enhances ER signaling and confers tamoxifen resistance in breast cancer. Oncogene 2016, 35, 2746–2755. [Google Scholar] [CrossRef]
- Borges, S.; Döppler, H.; Perez, E.A.; Andorfer, C.A.; Sun, Z.; Anastasiadis, P.Z.; Thompson, E.A.; Geiger, X.J.; Storz, P. Pharmacologic reversion of epigenetic silencing of the PRKD1 promoter blocks breast tumor cell invasion and metastasis. Breast Cancer Res. 2013, 15, R66. [Google Scholar] [CrossRef]
- Yu, J.; Qin, B.; Moyer, A.M.; Nowsheen, S.; Liu, T.; Qin, S.; Zhuang, Y.; Liu, D.; Lu, S.W.; Kalari, K.R.; et al. DNA methyltransferase expression in triple-negative breast cancer predicts sensitivity to decitabine. J. Clin. Invest. 2018, 128, 2376–2388. [Google Scholar] [CrossRef] [Green Version]
- Pitta, C.A.; Papageorgis, P.; Charalambous, C.; Constantinou, A.I. Reversal of ER-β silencing by chromatin modifying agents overrides acquired tamoxifen resistance. Cancer Lett. 2013, 337, 167–176. [Google Scholar] [CrossRef]
- Zhu, Y.; Shao, S.; Pan, H.; Cheng, Z.; Rui, X. MicroRNA-136 inhibits prostate cancer cell proliferation and invasion by directly targeting mitogen-activated protein kinase kinase 4. Mol. Med. Rep. 2018, 17, 4803–4810. [Google Scholar] [CrossRef]
- Li, D.-X.; Fei, X.-R.; Dong, Y.-F.; Cheng, C.-D.; Yang, Y.; Deng, X.-F.; Huang, H.-L.; Niu, W.-X.; Zhou, C.-X.; Xia, C.-Y.; et al. The long non-coding RNA CRNDE acts as a ceRNA and promotes glioma malignancy by preventing miR-136-5p-mediated downregulation of Bcl-2 and Wnt2. Oncotarget 2017, 8, 88163–88178. [Google Scholar] [CrossRef] [Green Version]
- Singh, A.; Willems, E.; Singh, A.; Ong, I.M.; Verma, A.K. Ultraviolet radiation-induced differential microRNA expression in the skin of hairless SKH1 mice, a widely used mouse model for dermatology research. Oncotarget 2016, 7, 84924–84937. [Google Scholar] [CrossRef] [Green Version]
- Wallacides, A.; Chesnel, A.; Ajj, H.; Chillet, M.; Flament, S.; Dumond, H. Estrogens promote proliferation of the seminoma-like TCam-2 cell line through a GPER-dependent ERα36 induction. Mol. Cell. Endocrinol. 2012, 350, 61–71. [Google Scholar] [CrossRef]
- Iorio, M.V.; Ferracin, M.; Liu, C.-G.; Veronese, A.; Spizzo, R.; Sabbioni, S.; Magri, E.; Pedriali, M.; Fabbri, M.; Campiglio, M.; et al. MicroRNA gene expression deregulation in human breast cancer. Cancer Res. 2005, 65, 7065–7070. [Google Scholar] [CrossRef]
- Paszek, S.; Gabło, N.; Barnaś, E.; Szybka, M.; Morawiec, J.; Kołacińska, A.; Zawlik, I. Dysregulation of microRNAs in triple-negative breast cancer. Ginekol. Pol. 2017, 88, 530–536. [Google Scholar] [CrossRef] [Green Version]
- Braicu, C.; Raduly, L.; Morar-Bolba, G.; Cojocneanu, R.; Jurj, A.; Pop, L.-A.; Pileczki, V.; Ciocan, C.; Moldovan, A.; Irimie, A.; et al. Aberrant miRNAs expressed in HER-2 negative breast cancers patient. J. Exp. Clin. Cancer Res. 2018, 37, 257. [Google Scholar] [CrossRef]
- Yerukala Sathipati, S.; Ho, S.-Y. Identifying a miRNA signature for predicting the stage of breast cancer. Sci. Rep. 2018, 8, 16138. [Google Scholar] [CrossRef]
- Cheng, D.; He, H.; Liang, B. A three-microRNA signature predicts clinical outcome in breast cancer patients. Eur. Rev. Med. Pharmacol. Sci. 2018, 22, 6386–6395. [Google Scholar]
- Kagami, M.; O’Sullivan, M.J.; Green, A.J.; Watabe, Y.; Arisaka, O.; Masawa, N.; Matsuoka, K.; Fukami, M.; Matsubara, K.; Kato, F.; Ferguson-Smith, A.C.; Ogata, T. The IG-DMR and the MEG3-DMR at human chromosome 14q32.2: Hierarchical interaction and distinct functional properties as imprinting control centers. PLoS Genet. 2010, 6, e1000992. [Google Scholar] [CrossRef]
- Zhu, L.; Liu, Y.; Chen, Q.; Yu, G.; Chen, J.; Chen, K.; Yang, N.; Zeng, T.; Yan, S.; Huang, A.; et al. Long-Noncoding RNA Colorectal Neoplasia Differentially Expressed Gene as a Potential Target to Upregulate the Expression of IRX5 by miR-136-5P to Promote Oncogenic Properties in Hepatocellular Carcinoma. Cell. Physiol. Biochem. 2018, 50, 2229–2248. [Google Scholar] [CrossRef]
- Li, B.; Wang, F.; Li, X.; Sun, S.; Shen, Y.; Yang, H. Hsa_circ_0008309 May Be a Potential Biomarker for Oral Squamous Cell Carcinoma. Dis. Markers 2018, 2018, 7496890. [Google Scholar] [CrossRef]
- Duan, C.; Liu, Y.; Li, Y.; Chen, H.; Liu, X.; Chen, X.; Yue, J.; Zhou, X.; Yang, J. Sulfasalazine alters microglia phenotype by competing endogenous RNA effect of miR-136-5p and long non-coding RNA HOTAIR in cuprizone-induced demyelination. Biochem. Pharmacol. 2018, 155, 110–123. [Google Scholar] [CrossRef]
- Dai, M.; Li, S.; Qin, X. Colorectal neoplasia differentially expressed: A long noncoding RNA with an imperative role in cancer. OncoTargets Ther. 2018, 11, 3755–3763. [Google Scholar] [CrossRef]
- Wang, W.; Yuan, F.; Xu, J. The prognostic role of long noncoding RNA CRNDE in cancer patients: A systematic review and meta-analysis. Neoplasma 2019, 66, 73–82. [Google Scholar] [CrossRef]
- Lin, C.-C.; Liu, L.-Z.; Addison, J.B.; Wonderlin, W.F.; Ivanov, A.V.; Ruppert, J.M. A KLF4-miRNA-206 autoregulatory feedback loop can promote or inhibit protein translation depending upon cell context. Mol. Cell. Biol. 2011, 31, 2513–2527. [Google Scholar] [CrossRef]
- Sobin, L.H.; Wittekind, C.H. TNM classification of malignant tumors, 6th ed; John Wiley & Sons: Hoboken, NJ, USA, 2002. [Google Scholar]
- Altman, D.G.; Bland, J.M. Standard deviations and standard errors. BMJ 2005, 331, 903. [Google Scholar] [CrossRef]
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Thiebaut, C.; Chesnel, A.; Merlin, J.-L.; Chesnel, M.; Leroux, A.; Harlé, A.; Dumond, H. Dual Epigenetic Regulation of ERα36 Expression in Breast Cancer Cells. Int. J. Mol. Sci. 2019, 20, 2637. https://doi.org/10.3390/ijms20112637
Thiebaut C, Chesnel A, Merlin J-L, Chesnel M, Leroux A, Harlé A, Dumond H. Dual Epigenetic Regulation of ERα36 Expression in Breast Cancer Cells. International Journal of Molecular Sciences. 2019; 20(11):2637. https://doi.org/10.3390/ijms20112637
Chicago/Turabian StyleThiebaut, Charlène, Amand Chesnel, Jean-Louis Merlin, Maelle Chesnel, Agnès Leroux, Alexandre Harlé, and Hélène Dumond. 2019. "Dual Epigenetic Regulation of ERα36 Expression in Breast Cancer Cells" International Journal of Molecular Sciences 20, no. 11: 2637. https://doi.org/10.3390/ijms20112637