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Licensed Unlicensed Requires Authentication Published by De Gruyter January 15, 2021

Vorinostat exhibits anticancer effects in triple-negative breast cancer cells by preventing nitric oxide-driven histone deacetylation

  • Marianne B. Palczewski , Hannah Petraitis Kuschman , Rhea Bovee , Jason R. Hickok and Douglas D. Thomas ORCID logo EMAIL logo
From the journal Biological Chemistry


Triple-negative breast cancers (TNBC) that produce nitric oxide (NO) are more aggressive, and the expression of the inducible form of nitric oxide synthase (NOS2) is a negative prognostic indicator. In these studies, we set out to investigate potential therapeutic strategies to counter the tumor-permissive properties of NO. We found that exposure to NO increased proliferation of TNBC cells and that treatment with the histone deacetylase inhibitor Vorinostat (SAHA) prevented this proliferation. When histone acetylation was measured in response to NO and/or SAHA, NO significantly decreased acetylation on histone 3 lysine 9 (H3K9ac) and SAHA increased H3K9ac. If NO and SAHA were sequentially administered to cells (in either order), an increase in acetylation was observed in all cases. Mechanistic studies suggest that the “deacetylase” activity of NO does not involve S-nitrosothiols or soluble guanylyl cyclase activation. The observed decrease in histone acetylation by NO required the interaction of NO with cellular iron pools and may be an overriding effect of NO-mediated increases in histone methylation at the same lysine residues. Our data revealed a novel pathway interaction of Vorinostat and provides new insight in therapeutic strategy for aggressive TNBCs.

Corresponding author: Douglas D. Thomas, Department of Pharmaceutical Sciences, University of Illinois at Chicago, 833 S. Wood Street, Chicago, IL60607, USA, E-mail: .
Marianne B. Palczewski and Hannah Petraitis Kuschman contributed equally to this work.


This research was supported in part by “bridge funding” from the UICancer Center.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: This research was supported in part by “bridge funding” from the UICancer Center.

  3. Conflict of interest statement: The authors declare no conflicts of interest regarding this article.


Bubna, A.K. (2015). Vorinostat-an overview. Indian J. Dermatol. 60: 419, in Google Scholar

De Paepe, B., Verstraeten, V.M., De Potter, C.R., and Bullock, G.R. (2002). Increased angiotensin II type-2 receptor density in hyperplasia, DCIS and invasive carcinoma of the breast is paralleled with increased iNOS expression. Histochem. Cell Biol. 117: 13–19, in Google Scholar

Dumitrescu, R.G. (2018). Interplay between genetic and epigenetic changes in breast cancer subtypes. Methods Mol. Biol. 1856: 19–34, in Google Scholar

Garmpis, N., Damaskos, C., Garmpi, A., Kalampokas, E., Kalampokas, T., Spartalis, E., Daskalopoulou, A., Valsami, S., Kontos, M., Nonni, A., et al.. (2017). Histone deacetylases as new therapeutic targets in triple-negative breast cancer: progress and promises. Cancer Genomics Proteomics 14: 299–313, in Google Scholar

Glynn, S.A., Boersma, B.J., Dorsey, T.H., Yi, M., Yfantis, H.G., Ridnour, L.A., Martin, D.N., Switzer, C.H., Hudson, R.S., Wink, D.A., et al.. (2010). Increased NOS2 predicts poor survival in estrogen receptor-negative breast cancer patients. J. Clin. Invest. 120: 3843–3854, in Google Scholar

Griffith, D.M., Szocs, B., Keogh, T., Suponitsky, K.Y., Farkas, E., Buglyo, P., and Marmion, C.J. (2011). Suberoylanilide hydroxamic acid, a potent histone deacetylase inhibitor; its X-ray crystal structure and solid state and solution studies of its Zn(II), Ni(II), Cu(II) and Fe(III) complexes. J. Inorg. Biochem. 105: 763–769, in Google Scholar

Hebbel, R.P., Vercellotti, G.M., Pace, B.S., Solovey, A.N., Kollander, R., Abanonu, C.F., Nguyen, J., Vineyard, J.V., Belcher, J.D., Abdulla, F., et al.. (2010). The HDAC inhibitors trichostatin A and suberoylanilide hydroxamic acid exhibit multiple modalities of benefit for the vascular pathobiology of sickle transgenic mice. Blood 115: 2483–2490, in Google Scholar

Heinecke, J.L., Ridnour, L.A., Cheng, R.Y., Switzer, C.H., Lizardo, M.M., Khanna, C., Glynn, S.A., Hussain, S.P., Young, H.A., Ambs, S., et al.. (2014). Tumor microenvironment-based feed-forward regulation of NOS2 in breast cancer progression. Proc. Natl. Acad. Sci. U.S.A. 111: 6323–6328, in Google Scholar

Hickok, J.R., Sahni, S., Mikhed, Y., Bonini, M.G., and Thomas, D.D. (2011a). Nitric oxide suppresses tumor cell migration through N-Myc downstream-regulated gene-1 (NDRG1) expression: role of chelatable iron. J. Biol. Chem. 286: 41413–41424, in Google Scholar

Hickok, J.R., Sahni, S., Shen, H., Arvind, A., Antoniou, C., Fung, L.W., and Thomas, D.D. (2011b). Dinitrosyliron complexes are the most abundant nitric oxide-derived cellular adduct: biological parameters of assembly and disappearance. Free Radic. Biol. Med. 51: 1558–1566, in Google Scholar

Hickok, J.R. and Thomas, D.D. (2010). Nitric oxide and cancer therapy: the emperor has NO clothes. Curr. Pharmaceut. Des. 16: 381–391, in Google Scholar

Hickok, J.R., Vasudevan, D., Antholine, W.E., and Thomas, D.D. (2013). Nitric oxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases. J. Biol. Chem. 288: 16004–16015, in Google Scholar

Hickok, J.R., Vasudevan, D., Thatcher, G.R., and Thomas, D.D. (2012). Is S-nitrosocysteine a true surrogate for nitric oxide? Antioxidants Redox Signal. 17: 962–968, in Google Scholar

Kroncke, K.D. (2001). Zinc finger proteins as molecular targets for nitric oxide-mediated gene regulation. Antioxidants Redox Signal. 3: 565–575, in Google Scholar

Loibl, S., Buck, A., Strank, C., von Minckwitz, G., Roller, M., Sinn, H.P., Schini-Kerth, V., Solbach, C., Strebhardt, K., and Kaufmann, M. (2005). The role of early expression of inducible nitric oxide synthase in human breast cancer. Eur. J. Canc. 41: 265–271. in Google Scholar

MacArthur, P.H., Shiva, S., and Gladwin, M.T. (2007). Measurement of circulating nitrite and S-nitrosothiols by reductive chemiluminescence. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 851: 93–105, in Google Scholar

Mendes, R.V., Martins, A.R., de Nucci, G., Murad, F., and Soares, F.A. (2001). Expression of nitric oxide synthase isoforms and nitrotyrosine immunoreactivity by B-cell non-Hodgkin’s lymphomas and multiple myeloma. Histopathology 39: 172–178, in Google Scholar

Munoz-Sanchez, J. and Chanez-Cardenas, M.E. (2019). The use of cobalt chloride as a chemical hypoxia model. J. Appl. Toxicol. 39: 556–570, in Google Scholar

Nakayama, J., Rice, J.C., Strahl, B.D., Allis, C.D., and Grewal, S.I. (2001). Role of histone H3 lysine 9 methylation in epigenetic control of heterochromatin assembly. Science 292: 110–113, in Google Scholar

Prueitt, R.L., Boersma, B.J., Howe, T.M., Goodman, J.E., Thomas, D.D., Ying, L., Pfiester, C.M., Yfantis, H.G., Cottrell, J.R., Lee, D.H., Remaley, A.T., Hofseth, L.J., Wink, D.A., and Ambs, S. (2007). Inflammation and IGF-I activate the Akt pathway in breast cancer. Int. J. Cancer 120: 796–805, in Google Scholar

Rao, C.V., Indranie, C., Simi, B., Manning, P.T., Connor, J.R., and Reddy, B.S. (2002). Chemopreventive properties of a selective inducible nitric oxide synthase inhibitor in colon carcinogenesis, administered alone or in combination with celecoxib, a selective cyclooxygenase-2 inhibitor. Cancer Res. 62: 165–170.Search in Google Scholar

Ridnour, L.A., Barasch, K.M., Windhausen, A.N., Dorsey, T.H., Lizardo, M.M., Yfantis, H.G., Lee, D.H., Switzer, C.H., Cheng, R.Y., Heinecke, J.L., et al.. (2012). Nitric oxide synthase and breast cancer: role of TIMP-1 in NO-mediated Akt activation. PloS One 7: e44081, in Google Scholar

Sabari, B.R., Zhang, D., Allis, C.D., and Zhao, Y. (2017). Metabolic regulation of gene expression through histone acylations. Nat. Rev. Mol. Cell Biol. 18: 90–101, in Google Scholar

Sahni, S., Hickok, J.R., and Thomas, D.D. (2018). Nitric oxide reduces oxidative stress in cancer cells by forming dinitrosyliron complexes. Nitric Oxide 76: 37–44, in Google Scholar

Shiva, S., Brookes, P.S., Patel, R.P., Anderson, P.G., and Darley-Usmar, V.M. (2001). Nitric oxide partitioning into mitochondrial membranes and the control of respiration at cytochrome c oxidase. Proc. Natl. Acad. Sci. U.S.A. 98: 7212–7217, in Google Scholar

Socco, S., Bovee, R.C., Palczewski, M.B., Hickok, J.R., and Thomas, D.D. (2017). Epigenetics: the third pillar of nitric oxide signaling. Pharmacol. Res. 121: 52–58, in Google Scholar

Switzer, C.H., Cheng, R.Y., Ridnour, L.A., Glynn, S.A., Ambs, S., and Wink, D.A. (2012a). Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer. Breast Cancer Res. 14: R125, in Google Scholar

Switzer, C.H., Glynn, S.A., Ridnour, L.A., Cheng, R.Y., Vitek, M.P., Ambs, S., and Wink, D.A. (2011). Nitric oxide and protein phosphatase 2A provide novel therapeutic opportunities in ER-negative breast cancer. Trends Pharmacol. Sci. 32: 644–651, in Google Scholar

Switzer, C.H., Ridnour, L.A., Cheng, R., Heinecke, J., Burke, A., Glynn, S., Ambs, S., and Wink, D.A. (2012b). S-Nitrosation mediates multiple pathways that lead to tumor progression in estrogen receptor-negative breast cancer. For. Immunopathol. Dis. Therap. 3: 117–124, in Google Scholar

Thomas, D.D. (2015). Breathing new life into nitric oxide signaling: a brief overview of the interplay between oxygen and nitric oxide. Redox Biol. 5: 225–233, in Google Scholar

Thomas, D.D., Heinecke, J.L., Ridnour, L.A., Cheng, R.Y., Kesarwala, A.H., Switzer, C.H., McVicar, D.W., Roberts, D.D., Glynn, S., Fukuto, J.M., et al.. (2015). Signaling and stress: the redox landscape in NOS2 biology. Free Radic. Biol. Med. 87: 204–225, in Google Scholar

Vasudevan, D., Bovee, R.C., and Thomas, D.D. (2016). Nitric oxide, the new architect of epigenetic landscapes. Nitric Oxide 59: 54–62, in Google Scholar

Vasudevan, D., Hickok, J.R., Bovee, R.C., Pham, V., Mantell, L.L., Bahroos, N., Kanabar, P., Cao, X.J., Maienschein-Cline, M., Garcia, B.A., and Thomas, D.D. (2015). Nitric oxide regulates gene expression in cancers by controlling histone posttranslational modifications. Cancer Res. 75: 5299–5308, in Google Scholar

Vasudevan, D. and Thomas, D.D. (2014). Insights into the diverse effects of nitric oxide on tumor biology. Vitam. Horm. 96: 265–298, in Google Scholar

Zhang, L., Zhang, J., Jiang, Q., Zhang, L., and Song, W. (2018). Zinc binding groups for histone deacetylase inhibitors. J. Enzyme Inhib. Med. Chem. 33: 714–721, in Google Scholar

Zhang, Y. and Reinberg, D. (2001). Transcription regulation by histone methylation: interplay between different covalent modifications of the core histone tails. Genes Dev. 15: 2343–2360, in Google Scholar

Zhao, L., Okhovat, J.P., Hong, E.K., Kim, Y.H., and Wood, G.S. (2019). Preclinical studies support combined inhibition of BET family proteins and histone deacetylases as epigenetic therapy for cutaneous T-cell lymphoma. Neoplasia 21: 82–92, in Google Scholar

Received: 2020-09-23
Accepted: 2020-12-18
Published Online: 2021-01-15
Published in Print: 2021-03-26

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