Home Quercetin increases mitochondrial proteins (VDAC and SDH) and downmodulates AXL and PIM-1 tyrosine kinase receptors in NRAS melanoma cells
Article
Licensed
Unlicensed Requires Authentication

Quercetin increases mitochondrial proteins (VDAC and SDH) and downmodulates AXL and PIM-1 tyrosine kinase receptors in NRAS melanoma cells

  • Karin J. P. Rocha-Brito , Stefano Piatto Clerici ORCID logo , Helon Guimarães Cordeiro ORCID logo , Amanda Petrina Scotá Ferreira , Emanuella Maria Barreto Fonseca ORCID logo , Paola R. Gonçalves , Júlia Laura F. Abrantes , Renato Milani , Renato Ramos Massaro , Silvya Stuchi Maria-Engler and Carmen V. Ferreira-Halder ORCID logo EMAIL logo
Published/Copyright: December 3, 2021
Biological Chemistry
From the journal Biological Chemistry Volume 403 Issue 3

Abstract

Melanoma is a type of skin cancer with low survival rates after it has metastasized. In order to find molecular differences that could represent targets of quercetin in anti-melanoma activity, we have chosen SKMEL-103 and SKMEL-28 melanoma cells and human melanocytes as models. Firstly, we observed that quercetin was able in reducing SKMEL-103 cell viability, but not in SKMEL-28. Besides that, quercetin treatment caused inhibition of AXL in both cell lines, but upregulation of PIM-1 in SKMEL-28 and downregulation in SKMEL-103. Moreover, HIF-1 alpha expression decreased in both cell lines. Interestingly, quercetin was more effective against SKMEL-103 than kinases inhibitors, such as Imatinib, Temsirolimus, U0126, and Erlotinib. Interestingly, we observed that while the levels of succinate dehydrogenase and voltage-dependent anion channel increased in SKMEL-103, both proteins were downregulated in SKMEL-28 after quercetin’s treatment. Furthermore, AKT, AXL, PIM-1, ABL kinases were much more active and chaperones HSP90, HSP70 and GAPDH were highly expressed in SKMEL-103 cells in comparison with melanocytes. Our findings indicate, for the first time, that the efficacy of quercetin to kill melanoma cells depends on its ability in inhibiting tyrosine kinase and upregulating mitochondrial proteins, at least when SKMEL-103 and SKMEL-28 cells response were compared.

Keywords: melanoma; mitochondria; protein kinases; quercetin
Corresponding author: Carmen V. Ferreira-Halder, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, Campinas 13083-862, SP, Brazil, E-mail:

Funding source: Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)

Award Identifier / Grant number: 303330/2013-9; 472321/2012-9; 141723/2019-0

Funding source: The São Paulo Research Foundation (FAPESP)

Award Identifier / Grant number: 2015/20412-7; 2018/03593-6

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

  2. Research funding: This study was supported by São Paulo Research Foundation (FAPESP) and The National Council for Scientific and Technological Development. H.G.C., P.R.G., C.V.F-H. and S.S.M-E. were supported by research fellowships from CNPq (141723/2019-0, 472321/2012-9, 303330/2013-9 and 304339/2017-2, respectively). C.V.F-H. and S.S.M-E. thanks FAPESP for sponsorship of the projects (2015/20412-7; 2017/04926-6). S.P.C. thanks FAPESP for the scholarship (2018/03593-6).

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

References

Alonso-Curbelo, D., Riveiro-Falkenbach, E., Pérez-Guijarro, E., Cifdaloz, M., Karras, P., Osterloh, L., Megías, D., Cañón, E., Calvo, T.G., Olmeda, D., et al.. (2014). RAB7 controls melanoma progression by exploiting a lineage-specific wiring of the endolysosomal pathway. Cancer Cell 26: 61–76. https://doi.org/10.1016/j.ccr.2014.04.030.Search in Google Scholar PubMed

Avagliano, A., Fiume, G., Pelagalli, A., Sanità, G., Ruocco, M.R., Montagnani, S., and Arcucci, A. (2020). Metabolic plasticity of melanoma cells and their crosstalk with tumor microenvironment. Front. Oncol. 10: 722. https://doi.org/10.3389/fonc.2020.00722.Search in Google Scholar PubMed PubMed Central

Bachmann, M. and Möröy, T. (2005). The serine/threonine kinase Pim-1. Int. J. Biochem. Cell Biol. 37: 726–730. https://doi.org/10.1016/j.biocel.2004.11.005.Search in Google Scholar PubMed

Bettum, I.J., Gorad, S.S., Barkovskaya, A., Pettersen, S., Moestue, S.A., Vasiliauskaite, K., Tenstad, E., Øyjord, T., Risa, Ø., Nygaard, V., et al.. (2015). Metabolic reprogramming supports the invasive phenotype in malignant melanoma. Cancer Lett. 366: 71–83. https://doi.org/10.1016/j.canlet.2015.06.006.Search in Google Scholar PubMed

Buss, G.D., Constantin, J., De Lima, L.C.N., Teodoro, G.R., Comar, J.F., Ishii-Iwamoto, E.L., and Bracht, A. (2005). The action of quercetin on the mitochondrial NADH to NAD+ ratio in the isolated perfused rat liver. Planta Med. 71: 1118–1122. https://doi.org/10.1055/s-2005-873174.Search in Google Scholar PubMed

Capello, M., Ferri-Borgogno, S., Cappello, P., and Novelli, F. (2011). Alpha-enolase: a promising therapeutic and diagnostic tumor target. FEBS J. 278: 1064–1074. https://doi.org/10.1111/j.1742-4658.2011.08025.x.Search in Google Scholar PubMed

Cornford, P.A., Dodson, A.R., Parsons, K.F., Desmond, A.D., Woolfenden, A., Fordham, M., Neoptolemos, J.P., Ke, Y., and Foster, C.S. (2000). Heat shock protein expression independently predicts clinical outcome in prostate cancer. Tumor Biol. 60: 7099–7105.Search in Google Scholar

Devipriya, S., Ganapathy, V., and Shyamaladevi, C.S. (2006). Suppression of tumor growth and invasion in 9,10 dimethyl benz(a) anthracene induced mammary carcinoma by the plant bioflavonoid quercetin. Chem. Biol. Interact. 162: 106–113. https://doi.org/10.1016/j.cbi.2006.04.002.Search in Google Scholar PubMed

Devitt, B., Liu, W., Salemi, R., Wolfe, R., Kelly, J., Tzen, C.Y., Dobrovic, A., and McArthur, G. (2011). Clinical outcome and pathological features associated with NRAS mutation in cutaneous melanoma. Pigment Cell Melanoma Res. 24: 666–672. https://doi.org/10.1111/j.1755-148x.2011.00873.x.Search in Google Scholar

Easty, D.J. and Bennett, D.C. (2000). Protein tyrosine kinases in malignant melanoma. Melanoma Res. 10: 401–411. https://doi.org/10.1097/00008390-200010000-00001.Search in Google Scholar PubMed

Easty, D.J., Gray, S.G., O’Byrne, K.J., O’Donnell, D., and Bennett, D.C. (2011). Receptor tyrosine kinases and their activation in melanoma. Pigment Cell Melanoma Res. 24: 446–461. https://doi.org/10.1111/j.1755-148x.2011.00836.x.Search in Google Scholar PubMed

Eskandarpour, M., Huang, F., Reeves, K.A., Clark, E.A., and Hansson, J. (2009). Oncogenic NRAS has multiple effects on the malignant phenotype of human melanoma cells cultured in vitro. Int. J. Cancer 124: 16–26. https://doi.org/10.1002/ijc.23876.Search in Google Scholar PubMed

Ferry, D.R., Smith, A., Malkhandi, J., Fyfe, D.W., deTakats, P.G., Anderson, D., Baker, J., and Kerr, D.J. (1996). Phase I clinical trial of the flavonoid quercetin: pharmacokinetics and evidence for in vivo tyrosine kinase inhibition. Clin. Cancer Res. 2: 659–668.Search in Google Scholar

Fiorani, M., Guidarelli, A., Blasa, M., Azzolini, C., Candiracci, M., Piatti, E., and Cantoni, O. (2010). Mitochondria accumulate large amounts of quercetin: prevention of mitochondrial damage and release upon oxidation of the extramitochondrial fraction of the flavonoid. J. Nutr. Biochem. 21: 397–404. https://doi.org/10.1016/j.jnutbio.2009.01.014.Search in Google Scholar PubMed

Funakoshi, T., Kanzaki, N., Otsuka, Y., Izumo, T., Shibata, H., and Machida, S. (2017). Quercetin inhibits adipogenesis of muscle progenitor cells in vitro. Biochem. Biophys. Rep. 13: 39–44. https://doi.org/10.1016/j.bbrep.2017.12.003.Search in Google Scholar PubMed PubMed Central

Garbe, C., Eigentler, T.K., Keilholz, U., Hauschild, A., and Kirkwood, J.M. (2011). Systematic review of medical treatment in melanoma: current status and future prospects. Oncologist. 16: 5–24. https://doi.org/10.1634/theoncologist.2010-0190.Search in Google Scholar PubMed PubMed Central

Gibellini, L., Pinti, M., Nasi, M., Montagna, J.P., De Biasi, S., Roat, E., Bertoncelli, L., Cooper, E.L., and Cossarizza, A. (2011). Quercetin and cancer chemoprevention. Evid. Based Complement. Altern. Med. 2011: 591356. https://doi.org/10.1093/ecam/neq053.Search in Google Scholar PubMed PubMed Central

Gorlach, S., Fichna, J., and Lewandowska, U. (2015). Polyphenols as mitochondria-targeted anticancer drugs. Cancer Lett. 366: 141–149. https://doi.org/10.1016/j.canlet.2015.07.004.Search in Google Scholar PubMed

Greger, J.G., Eastman, S.D., Zhang, V., Bleam, M.R., Hughes, A.M. and Smitheman, K.N. (2012). Combinations of BRAF, MEK, and PI3K/mTOR inhibitors overcome acquired resistance to the BRAF inhibitor GSK2118436 dabrafenib, mediated by NRAS or MEK mutations. Mol. Cancer Ther. 11: 909–920. https://doi.org/10.1158/1535-7163.mct-11-0989.Search in Google Scholar PubMed

Ippolito, L., Marini, A., Cavallini, L., Morandi, A., Pietrovito, L., Pintus, G., Giannoni, E., Schrader, T., Puhr, M., Chiarugi, P., et al.. (2016). Metabolic shift toward oxidative phosphorylation in docetaxel resistant prostate cancer cells. Oncotarget 5: 61890–61904. https://doi.org/10.18632/oncotarget.11301.Search in Google Scholar PubMed PubMed Central

Johnson, D.B., Menzies, A.M., Zimmer, L., Eroglu, Z., Ye, F., and Zhao, S. (2015). Acquired BRAF inhibitor resistance: a multicenter meta-analysis of the spectrum and frequencies, clinical behavior, and phenotypic associations of resistance mechanisms. Eur. J. Cancer 51: 2792–2799. https://doi.org/10.1016/j.ejca.2015.08.022.Search in Google Scholar PubMed PubMed Central

Johnson, G.L., Stuhlmiller, T.J., Angus, S.P., Zawistowski, J.S., and Graves, L.M. (2014). Molecular pathways: adaptive kinome reprogramming in response to targeted inhibition of the BRAF-MEK-ERK pathway in cancer. Clin. Cancer Res. 20: 2516–2522. https://doi.org/10.1158/1078-0432.ccr-13-1081.Search in Google Scholar

Kardos, G.R., Dai, M.S., and Robertson, G.P. (2014). Growth inhibitory effects of large subunit ribosomal proteins in melanoma. Pigment Cell Melanoma Res. 27: 801–812. https://doi.org/10.1111/pcmr.12259.Search in Google Scholar PubMed PubMed Central

Le, B.T., Kumarasiri, M., Adams, J.R., Yu, M., Milne, R., Sykes, M.J., and Wang, S. (2015). Targeting Pim kinases for cancer treatment: opportunities and challenges. Future Med. Chem. 7: 35–53. https://doi.org/10.4155/fmc.14.145.Search in Google Scholar PubMed

Leiherer, A., Stoemmer, K., Muendlein, A., Saely, C.H., Kinz, E., Brandtner, E.M., Fraunberger, P., and Drexel, H. (2016). Quercetin impacts expression of metabolism-and obesity-associated genes in SGBS adipocytes. Nutrients 8: 282. https://doi.org/10.3390/nu8050282.Search in Google Scholar PubMed PubMed Central

Li, Y., Jia, L., Ren, D., Liu, C., Gong, Y., Wang, N., Zhang, X., and Zhao, Y. (2014). Axl mediates tumor invasion and chemosensitivity through PI3K/Akt signaling pathway and is transcriptionally regulated by slug in breast carcinoma. IUBMB Life 66: 507–518. https://doi.org/10.1002/iub.1285.Search in Google Scholar PubMed

Long, G.V., Menzies, A.M., Nagrial, A.M., Haydu, L.E., Hamilton, A.L., Mann, G.J., Hughes, T.M., Thompson, J.F., Scolyer, R.A., and Kefford, R.F. (2011). Prognostic and clinicopathologic associations of oncogenic BRAF in metastatic melanoma. J. Clin. Oncol. 29: 1239–1246. https://doi.org/10.1200/jco.2010.32.4327.Search in Google Scholar PubMed

Lopes, B.A., Ortelan, A.G., Faria, A.V.S., and Ferreira-Halder, C.V. (2020). Melanoma cell lines: mutations and impact on signal transduction pathways. Braz. J. Healthy Rev. 3: 10859–10884. https://doi.org/10.34119/bjhrv3n4-291.Search in Google Scholar

Mahalingam, D., Swords, R., Carew, J.S., Nawrocki, S.T., Bhalla, K., and Giles, F.J. (2009). Targeting HSP90 for cancer therapy. Br. J. Cancer 100: 1523–1529. https://doi.org/10.1038/sj.bjc.6605066.Search in Google Scholar PubMed PubMed Central

Miller, C.J. and Turk, B.E. (2018). Homing in: mechanisms of substrate targeting by protein kinases. Trends Biochem. Sci. 43: 380–394. https://doi.org/10.1016/j.tibs.2018.02.009.Search in Google Scholar PubMed PubMed Central

Mizuno, K., Shirogane, T., Shinohara, A., Iwamatsu, A., Hibi, M., and Hirano, T. (2001). Regulation of pim-1 by Hsp90. Biochem. Biophys. Res. Commun. 281: 663–669. https://doi.org/10.1006/bbrc.2001.4405.Search in Google Scholar PubMed

Mori, M., Vignaroli, G., and Botta, M. (2013). Small molecules modulation of 14-3-3 protein-protein interactions. Drug Discov. Today Technol. 10: e541–7. https://doi.org/10.1016/j.ddtec.2012.10.001.Search in Google Scholar PubMed

Morrison, D.K. (2009). The 14-3-3 proteins: integrators of diverse signaling cues that impact cell fate and cancer development. Trends Cell Biol. 19: 16–23. https://doi.org/10.1016/j.tcb.2008.10.003.Search in Google Scholar

Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65: 55–63. https://doi.org/10.1016/0022-1759(83)90303-4.Search in Google Scholar

Nazarian, R., Shi, H., Wang, Q., Kong, X., Koya, R.C., and Lee, H. (2010). Melanomas acquire resistance to B-RAF(V600E) inhibition by RTK or N-RAS upregulation. Nature 468: 973–977. https://doi.org/10.1038/nature09626.Search in Google Scholar PubMed PubMed Central

Neuzil, J., Dong, L.F., Rohlena, J., Truksa, J., and Ralph, S.J. (2013). Classification of mitocans, anti-cancer drugs acting on mitochondria. Mitochondrion. 13: 199–208. https://doi.org/10.1016/j.mito.2012.07.112.Search in Google Scholar PubMed

Nishimura, K., Kumazawa, T., Kuroda, T., Katagiri, N., Tsuchiya, M., Goto, N., Furumai, R., Murayama, A., Yanagisawa, J., and Kimura, K. (2015). Perturbation of ribosome biogenesis drives cells into senescence through 5S RNP-mediated p53 activation. Cell Rep. 10: 1310–1323. https://doi.org/10.1016/j.celrep.2015.01.055.Search in Google Scholar PubMed

Oji, Y., Tatsumi, N., Fukuda, M., Nakatsuka, S., Aoyagi, S., Hirata, E., Nanchi, I., Fujiki, F., Nakajima, H., Yamamoto, Y., et al.. (2014). The translation elongation factor eEF2 is a novel tumor-associated antigen overexpressed in various types of cancers. Int. J. Oncol. 44: 1461–1469. https://doi.org/10.3892/ijo.2014.2318.Search in Google Scholar PubMed PubMed Central

Pennacchi, P.C., de Almeida, M.E.S., Gomes, O.L.A., Faião-Flores, F., de Araújo Crepaldi, M.C., Dos Santos, M.F., de Moraes Barros, S.B., and Maria-Engler, S.S. (2015). Glycated reconstructed human skin as a platform to study the pathogenesis of skin aging. Tissue Eng. 21: 2417–2425. https://doi.org/10.1089/ten.tea.2015.0009.Search in Google Scholar PubMed

Penzo, M., Montanaro, L., Treré, D., and Derenzini, M. (2019). The ribosome biogenesis-cancer connection. Cells 8: 55. https://doi.org/10.3390/cells8010055.Search in Google Scholar PubMed PubMed Central

Porter, G.W., Khuri, F.R., and Fu, H. (2006). Dynamic 14-3-3/client protein interactions integrate survival and apoptotic pathways. Semin. Cancer Biol. 16: 193–202. https://doi.org/10.1016/j.semcancer.2006.03.003.Search in Google Scholar PubMed

Renzi, D., Valtolini, M., and Forster, R. (1993). The evaluation of a multi-endpoint cytotoxicity assay system. ATLA 21: 89–96. https://doi.org/10.1177/026119299302100114.Search in Google Scholar

Riddell, R.J., Clothier, R.H., and Balls, M. (1986). An evaluation of three in vitro cytotoxicity assays. Food Chem. Toxicol. 24: 469–471. https://doi.org/10.1016/0278-6915(86)90095-5.Search in Google Scholar

Ruggero, D. and Pandolfi, P.P. (2003). Does the ribosome translate cancer? Nat. Rev. Cancer. 3: 179–192. https://doi.org/10.1038/nrc1015.Search in Google Scholar

Sensi, M., Catani, M., Castellano, G., Nicolini, G., Alciato, F., Tragni, G., De Santis, G., Bersani, I., Avanzi, G., Tomassetti, A., et al.. (2011). Human cutaneous melanomas lacking MITF and melanocyte differentiation antigens express a functional Axl receptor kinase. J. Invest. Dermatol. 131: 2448–2457. https://doi.org/10.1038/jid.2011.218.Search in Google Scholar

Solit, D.B. and Chiosis, G. (2008). Development and application of Hsp90 inhibitors. Drug Discov. Today 13: 38–43. https://doi.org/10.1016/j.drudis.2007.10.007.Search in Google Scholar

Soll, F., Ternent, C., Berry, I.M., Kumari, D., and Moore, T.C. (2020). Quercetin inhibits proliferation and induces apoptosis of B16 melanoma cells in vitro. Assay Drug Dev. Technol. 18: 261–268. https://doi.org/10.1089/adt.2020.993.Search in Google Scholar

Wang, J.Y. (2000). Regulation of cell death by the Abl tyrosine kinase. Oncogene 19: 5643–5650. https://doi.org/10.1038/sj.onc.1203878.Search in Google Scholar

Warfel, N.A. and Kraft, A.S. (2015). PIM kinase (and Akt) biology and signaling in tumors. Pharmacol. Ther. 151: 41–49. https://doi.org/10.1016/j.pharmthera.2015.03.001.Search in Google Scholar

Xu, J.Q., Fan, N., Yu, B.Y., Wang, Q.Q., and Zhang, J. (2017). Biotransformation of quercetin by Gliocladium deliquescens NRRL 1086. Chin. J. Nat. Med. 15: 615–624. https://doi.org/10.1016/s1875-5364(17)30089-4.Search in Google Scholar

Yang, K., Oak, A., Slominski, R.M., Brożyna, A.A., and Slominski, A.T. (2020). Current molecular markers of melanoma and treatment targets. Int. J. Mol. Sci. 21: 3535. https://doi.org/10.3390/ijms21103535.Search in Google Scholar PubMed PubMed Central

Zhang, C., Yang, C., Wang, R., Jiao, Y., Ampah, K.K., Wang, X., and Zeng, X. (2013). c-Abl kinase is a regulator of alphavbeta3 integrin mediated melanoma A375 cell migration. PLoS ONE 8: e66108. https://doi.org/10.1371/journal.pone.0066108.Search in Google Scholar PubMed PubMed Central

Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/hsz-2021-0261).

Received: 2021-05-13
Accepted: 2021-11-18
Published Online: 2021-12-03
Published in Print: 2022-02-23

© 2021 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Research Articles
  3. Inhibitory effect of mitoquinone against the α-synuclein fibrillation and relevant neurotoxicity: possible role in inhibition of Parkinson’s disease
  4. Effect of myeloperoxidase oxidation and N-homocysteinylation of high-density lipoprotein on endothelial repair function
  5. High cellulose diet promotes intestinal motility through regulating intestinal immune homeostasis and serotonin biosynthesis
  6. Quercetin increases mitochondrial proteins (VDAC and SDH) and downmodulates AXL and PIM-1 tyrosine kinase receptors in NRAS melanoma cells
  7. Regulation of transforming growth factor-β1-stimulation of Runx2 acetylation for matrix metalloproteinase 13 expression in osteoblastic cells
  8. M6A methylation-mediated elevation of SM22α inhibits the proliferation and migration of vascular smooth muscle cells and ameliorates intimal hyperplasia in type 2 diabetes mellitus
  9. Characterization of hepatic zonation in mice by mass-spectrometric and antibody-based proteomics approaches
  10. Characterization of a fluorescent 1,8-naphthalimide-functionalized PAMAM dendrimer and its Cu(ii) complexes as cytotoxic drugs: EPR and biological studies in myeloid tumor cells
https://doi.org/10.1515/hsz-2021-0261
Keywords for this article
melanoma; mitochondria; protein kinases; quercetin

Articles in the same Issue

  1. Frontmatter
  2. Research Articles
  3. Inhibitory effect of mitoquinone against the α-synuclein fibrillation and relevant neurotoxicity: possible role in inhibition of Parkinson’s disease
  4. Effect of myeloperoxidase oxidation and N-homocysteinylation of high-density lipoprotein on endothelial repair function
  5. High cellulose diet promotes intestinal motility through regulating intestinal immune homeostasis and serotonin biosynthesis
  6. Quercetin increases mitochondrial proteins (VDAC and SDH) and downmodulates AXL and PIM-1 tyrosine kinase receptors in NRAS melanoma cells
  7. Regulation of transforming growth factor-β1-stimulation of Runx2 acetylation for matrix metalloproteinase 13 expression in osteoblastic cells
  8. M6A methylation-mediated elevation of SM22α inhibits the proliferation and migration of vascular smooth muscle cells and ameliorates intimal hyperplasia in type 2 diabetes mellitus
  9. Characterization of hepatic zonation in mice by mass-spectrometric and antibody-based proteomics approaches
  10. Characterization of a fluorescent 1,8-naphthalimide-functionalized PAMAM dendrimer and its Cu(ii) complexes as cytotoxic drugs: EPR and biological studies in myeloid tumor cells
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 18.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/hsz-2021-0261/html
Scroll to top button