Advertisement

Archives of Pharmacal Research

, Volume 42, Issue 6, pp 531–542 | Cite as

Flavonoid morin inhibits proliferation and induces apoptosis of melanoma cells by regulating reactive oxygen species, Sp1 and Mcl-1

  • Yoon Jin Lee
  • Woo Il Kim
  • Soo Young Kim
  • Sung Woo Cho
  • Hae Seon Nam
  • Sang Han Lee
  • Moon Kyun ChoEmail author
Research Article

Abstract

Reactive oxygen species (ROS) is associated with cancer progression in different cancers, including melanoma. It also affects specificity protein (Sp1), a transcription factor. Flavonoid morin is known to inhibit growth of cancer cells, including lung cancer and breast cancer. Herein, we hypothesized that morin can inhibit cancer activities in melanoma by altering ROS generation. The aim of this study is to determine the effects of morin and its underlying mechanisms in melanoma cells. Effects of morin on cell proliferation and apoptosis were determined using standardized assays. Changes in pro-apoptotic and anti-apoptotic proteins were analyzed by western blot analysis. Cellular ROS levels and mitochondrial function were evaluated by measuring DCF-DA fluorescence and rhodamine-123 fluorescence intensities, respectively. Morin induced ROS production and apoptosis, as presented by increased proportion of cells with Annexin V-PE(+) staining and sub-G0/G1 peak in cell cycle analysis. It also downregulated Sp1, Mcl-1, Bcl-2, and caspase-3 but upregulated cleaved caspase-3, Bax, and PUMA. In immunohistochemical staining, Sp1 was overexpressed in melanoma tissues compared to normal skin tissues. Collectively, our data suggest that morin can induce apoptosis of melanoma cells by regulating pro-apoptotic and anti-apoptotic proteins through ROS, and may be a potential substance for treatment of melanoma.

Keywords

Morin Melanoma Proliferation ROS Sp1 Mcl-1 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

References

  1. Arbour N, Vanderluit JL, Le Grand JN, Jahani-Asl A, Ruzhynsky VA, Cheung EC, Kelly MA, Mackenzie AE, Park DS, Opferman JT, Slack RS (2008) Mcl-1 is a key regulator of apoptosis during CNS development and after DNA damage. J Neurosci 28:6068–6078CrossRefGoogle Scholar
  2. Bajpai R, Nagaraju GP (2017) Specificity protein 1: its role in colorectal cancer progression and metastasis. Crit Rev Oncol Hematol 113:1–7CrossRefGoogle Scholar
  3. Baldelli S, Aquilano K, Rotilio G, Ciriolo MR (2008) Glutathione and copper, zinc superoxide dismutase are modulated by overexpression of neuronal nitric oxide synthase. Int J Biochem Cell Biol 40:2660–2670CrossRefGoogle Scholar
  4. Beroukhim R, Mermel CH, Porter D, Wei G, Raychaudhuri S, Donovan J, Barretina J, Boehm JS, Dobson J, Urashima M, Mc Henry KT, Pinchback RM, Ligon AH, Cho YJ, Haery L, Greulich H, Reich M, Winckler W, Lawrence MS, Weir BA, Tanaka KE, Chiang DY, Bass AJ, Loo A, Hoffman C, Prensner J, Liefeld T, Gao Q, Yecies D, Signoretti S, Maher E, Kaye FJ, Sasaki H, Tepper JE, Fletcher JA, Tabernero J, Baselga J, Tsao MS, Demichelis F, Rubin MA, Janne PA, Daly MJ, Nucera C, Levine RL, Ebert BL, Gabriel S, Rustgi AK, Antonescu CR, Ladanyi M, Letai A, Garraway LA, Loda M, Beer DG, True LD, Okamoto A, Pomeroy SL, Singer S, Golub TR, Lander ES, Getz G, Sellers WR, Meyerson M (2010) The landscape of somatic copy-number alteration across human cancers. Nature 463:899–905CrossRefGoogle Scholar
  5. Black AR, Black JD, Azizkhan-Clifford J (2001) Sp1 and kruppel-like factor family of transcription factors in cell growth regulation and cancer. J Cell Physiol 188:143–160CrossRefGoogle Scholar
  6. Briggs MR, Kadonaga JT, Bell SP, Tjian R (1986) Purification and biochemical characterization of the promoter-specific transcription factor, Sp1. Science 234:47–52CrossRefGoogle Scholar
  7. Brown J, O’prey J, Harrison PR (2003) Enhanced sensitivity of human oral tumours to the flavonol, morin, during cancer progression: involvement of the Akt and stress kinase pathways. Carcinogenesis 24:171–177CrossRefGoogle Scholar
  8. Cabello CM, Lamore SD, Bair WB 3rd, Qiao S, Azimian S, Lesson JL, Wondrak GT (2012) The redox antimalarial dihydroartemisinin targets human metastatic melanoma cells but not primary melanocytes with induction of NOXA-dependent apoptosis. Invest New Drugs 30:1289–1301CrossRefGoogle Scholar
  9. Chiefari E, Brunetti A, Arturi F, Bidart JM, Russo D, Schlumberger M, Filetti S (2002) Increased expression of AP2 and Sp1 transcription factors in human thyroid tumors: a role in NIS expression regulation? BMC Cancer 2:35CrossRefGoogle Scholar
  10. Choi ES, Han G, Park SK, Lee K, Kim HJ, Cho SD, Kim HM (2013) A248, a novel synthetic HDAC inhibitor, induces apoptosis through the inhibition of specificity protein 1 and its downstream proteins in human prostate cancer cells. Mol Med Rep 8:195–200CrossRefGoogle Scholar
  11. Choi CY, Kim JY, Wee SY, Lee JH, Nam DH, Kim CH, Cho MK, Lee YJ, Nam HS, Lee SH, Cho SW (2014) Expression of nuclear factor erythroid 2 protein in malignant cutaneous tumors. Arch Plast Surg 41:654–660CrossRefGoogle Scholar
  12. Chuang JY, Kao TJ, Lin SH, Wu AC, Lee PT, Su TP, Yeh SH, Lee YC, Wu CC, Chang WC (2017) Specificity protein 1-zinc finger protein 179 pathway is involved in the attenuation of oxidative stress following brain injury. Redox Biol 11:135–143CrossRefGoogle Scholar
  13. Deniaud E, Baguet J, Mathieu AL, Pages G, Marvel J, Leverrier Y (2006) Overexpression of Sp1 transcription factor induces apoptosis. Oncogene 25:7096–7105CrossRefGoogle Scholar
  14. Du Y, Qu J, Zhang W, Bai M, Zhou Q, Zhang Z, Li Z, Miao J (2016) Morin reverses neuropathological and cognitive impairments in APPswe/PS1dE9 mice by targeting multiple pathogenic mechanisms. Neuropharmacology 108:1–13CrossRefGoogle Scholar
  15. Duan H, Heckman CA, Boxer LM (2005) Histone deacetylase inhibitors down-regulate bcl-2 expression and induce apoptosis in t(14;18) lymphomas. Mol Cell Biol 25:1608–1619CrossRefGoogle Scholar
  16. Dzhagalov I, St John A, He YW (2007) The antiapoptotic protein Mcl-1 is essential for the survival of neutrophils but not macrophages. Blood 109:1620–1626CrossRefGoogle Scholar
  17. Dzhagalov I, Dunkle A, He YW (2008) The anti-apoptotic Bcl-2 family member Mcl-1 promotes T lymphocyte survival at multiple stages. J Immunol 181:521–528CrossRefGoogle Scholar
  18. Gandhy SU, Kim K, Larsen L, Rosengren RJ, Safe S (2012) Curcumin and synthetic analogs induce reactive oxygen species and decreases specificity protein (Sp) transcription factors by targeting microRNAs. BMC Cancer 12:564CrossRefGoogle Scholar
  19. Guan H, Cai J, Zhang N, Wu J, Yuan J, Li J, Li M (2012) Sp1 is upregulated in human glioma, promotes MMP-2-mediated cell invasion and predicts poor clinical outcome. Int J Cancer 130:593–601CrossRefGoogle Scholar
  20. Guo Z, Zhang W, Xia G, Niu L, Zhang Y, Wang X, Zhang Y, Jiang B, Wang J (2010) Sp1 upregulates the four and half lim 2 (FHL2) expression in gastrointestinal cancers through transcription regulation. Mol Carcinog 49:826–836Google Scholar
  21. Guterres FA, Martinez GR, Rocha ME, Winnischofer SM (2013) Simvastatin rises reactive oxygen species levels and induces senescence in human melanoma cells by activation of p53/p21 pathway. Exp Cell Res 319:2977–2988CrossRefGoogle Scholar
  22. Hambright HG, Meng P, Kumar AP, Ghosh R (2015) Inhibition of PI3 K/AKT/mTOR axis disrupts oxidative stress-mediated survival of melanoma cells. Oncotarget 6:7195–7208CrossRefGoogle Scholar
  23. Hofmann UB, Westphal JR, Van Muijen GN, Ruiter DJ (2000) Matrix metalloproteinases in human melanoma. J Invest Dermatol 115:337–344CrossRefGoogle Scholar
  24. Hsu TI, Wang MC, Chen SY, Yeh YM, Su WC, Chang WC, Hung JJ (2012) Sp1 expression regulates lung tumor progression. Oncogene 31:3973–3988CrossRefGoogle Scholar
  25. Hyun HB, Lee WS, Go SI, Nagappan A, Park C, Han MH, Hong SH, Kim G, Kim GY, Cheong J, Ryu CH, Shin SC, Choi YH (2015) The flavonoid morin from Moraceae induces apoptosis by modulation of Bcl-2 family members and Fas receptor in HCT 116 cells. Int J Oncol 46:2670–2678CrossRefGoogle Scholar
  26. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D (2011) Global cancer statistics. CA Cancer J Clin 61:69–90CrossRefGoogle Scholar
  27. Jiang NY, Woda BA, Banner BF, Whalen GF, Dresser KA, Lu D (2008) Sp1, a new biomarker that identifies a subset of aggressive pancreatic ductal adenocarcinoma. Cancer Epidemiol Biomark Prev 17:1648–1652CrossRefGoogle Scholar
  28. Jin M, Ande A, Kumar A, Kumar S (2013) Regulation of cytochrome P450 2e1 expression by ethanol: role of oxidative stress-mediated pkc/jnk/sp1 pathway. Cell Death Dis 4:e554CrossRefGoogle Scholar
  29. Jin H, Lee WS, Eun SY, Jung JH, Park HS, Kim G, Choi YH, Ryu CH, Jung JM, Hong SC, Shin SC, Kim HJ (2014) Morin, a flavonoid from Moraceae, suppresses growth and invasion of the highly metastatic breast cancer cell line MDA-MB231 partly through suppression of the Akt pathway. Int J Oncol 45:1629–1637CrossRefGoogle Scholar
  30. Karlseder J, Rotheneder H, Wintersberger E (1996) Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Mol Cell Biol 16:1659–1667CrossRefGoogle Scholar
  31. Kawabata K, Tanaka T, Honjo S, Kakumoto M, Hara A, Makita H, Tatematsu N, Ushida J, Tsuda H, Mori H (1999) Chemopreventive effect of dietary flavonoid morin on chemically induced rat tongue carcinogenesis. Int J Cancer 83:381–386CrossRefGoogle Scholar
  32. Kim HK, Namgoong SY, Kim HP (1993) Antiinflammatory activity of flavonoids: mouse ear edema inhibition. Arch Pharm Res 16:18CrossRefGoogle Scholar
  33. Klaunig JE, Xu Y, Bachowski S, Ketcham CA, Isenberg JS, Kolaja KL, Baker TK, Walborg EF Jr, Stevenson DE (1995) Oxidative stress in nongenotoxic carcinogenesis. Toxicol Lett 82–83:683–691CrossRefGoogle Scholar
  34. Kuo HM, Chang LS, Lin YL, Lu HF, Yang JS, Lee JH, Chung JG (2007) Morin inhibits the growth of human leukemia HL-60 cells via cell cycle arrest and induction of apoptosis through mitochondria dependent pathway. Anticancer Res 27:395–405Google Scholar
  35. Lau AT, Wang Y, Chiu JF (2008) Reactive oxygen species: current knowledge and applications in cancer research and therapeutic. J Cell Biochem 104:657–667CrossRefGoogle Scholar
  36. Lee SJ, Son KH, Chang HW, Do JC, Jung KY, Kang SS, Kim HP (1993) Antiinflammatory activity of naturally occurring flavone and flavonol glycosides. Arch Pharmacal Res 16:25CrossRefGoogle Scholar
  37. Lee HS, Jung K-H, Hong S-W, Park I-S, Lee C, Han H-K, Lee D-H, Hong S-S (2008) Morin protects acute liver damage by carbon tetrachloride (CCl4) in rat. Arch Pharm Res 31:1160–1165CrossRefGoogle Scholar
  38. Lee JH, Won YS, Park KH, Lee MK, Tachibana H, Yamada K, Seo KI (2012) Celastrol inhibits growth and induces apoptotic cell death in melanoma cells via the activation ROS-dependent mitochondrial pathway and the suppression of PI3 K/AKT signaling. Apoptosis 17:1275–1286CrossRefGoogle Scholar
  39. Lee J, Jin H, Lee WS, Nagappan A, Choi YH, Kim GS, Jung J, Ryu CH, Shin SC, Hong SC, Kim HJ (2016) Morin, a flavonoid from Moraceae, inhibits cancer cell adhesion to endothelial cells and EMT by downregulating VCAM1 and Ncadherin. Asian Pac J Cancer Prev 17:3071–3075Google Scholar
  40. Lee MH, Cha HJ, Choi EO, Han MH, Kim SO, Kim GY, Hong SH, Park C, Moon SK, Jeong SJ, Jeong MJ, Kim WJ, Choi YH (2017) Antioxidant and cytoprotective effects of morin against hydrogen peroxide-induced oxidative stress are associated with the induction of Nrf-2mediated HO-1 expression in V79-4 Chinese hamster lung fibroblasts. Int J Mol Med 39:672–680CrossRefGoogle Scholar
  41. Marnett LJ, Riggins JN, West JD (2003) Endogenous generation of reactive oxidants and electrophiles and their reactions with DNA and protein. J Clin Invest 111:583–593CrossRefGoogle Scholar
  42. Nguyen T, Nioi P, Pickett CB (2009) The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 284:13291–13295CrossRefGoogle Scholar
  43. Noor H, Cao P, Raleigh DP (2012) Morin hydrate inhibits amyloid formation by islet amyloid polypeptide and disaggregates amyloid fibers. Protein Sci 21:373–382CrossRefGoogle Scholar
  44. Ola MS, Aleisa AM, Al-Rejaie SS, Abuohashish HM, Parmar MY, Alhomida AS, Ahmed MM (2014) Flavonoid, morin inhibits oxidative stress, inflammation and enhances neurotrophic support in the brain of streptozotocin-induced diabetic rats. Neurol Sci 35:1003–1008CrossRefGoogle Scholar
  45. Opferman JT, Letai A, Beard C, Sorcinelli MD, Ong CC, Korsmeyer SJ (2003) Development and maintenance of B and T lymphocytes requires antiapoptotic MCL-1. Nature 426:671–676CrossRefGoogle Scholar
  46. Opferman JT, Iwasaki H, Ong CC, Suh H, Mizuno S, Akashi K, Korsmeyer SJ (2005) Obligate role of anti-apoptotic MCL-1 in the survival of hematopoietic stem cells. Science 307:1101–1104CrossRefGoogle Scholar
  47. Pervaiz S, Clement MV (2007) Superoxide anion: oncogenic reactive oxygen species? Int J Biochem Cell Biol 39:1297–1304CrossRefGoogle Scholar
  48. Prathyusha AMVN, Nawadkar R, Chari B (2017) Role of Sp1 Transcriptional Factor in Gastrointestinal Carcinogenesis. In: Nagaraju GP (ed) Role of transcription factors in gastrointestinal malignancies, 1st edn. Springer, Singapore, pp 191–201Google Scholar
  49. Reuter S, Gupta SC, Chaturvedi MM, Aggarwal BB (2010) Oxidative stress, inflammation, and cancer: how are they linked? Free Radic Biol Med 49:1603–1616CrossRefGoogle Scholar
  50. Sander CS, Hamm F, Elsner P, Thiele JJ (2003) Oxidative stress in malignant melanoma and non-melanoma skin cancer. Br J Dermatol 148:913–922CrossRefGoogle Scholar
  51. Sithara T, Arun KB, Syama HP, Reshmitha TR, Nisha P (2017) Morin inhibits proliferation of SW480 colorectal cancer cells by inducing apoptosis mediated by reactive oxygen species formation and uncoupling of warburg effect. Front Pharmacol 8:640CrossRefGoogle Scholar
  52. Sivaramakrishnan V, Niranjali Devaraj S (2009) Morin regulates the expression of NF-kappaB-p65, COX-2 and matrix metalloproteinases in diethylnitrosamine induced rat hepatocellular carcinoma. Chem Biol Interact 180:353–359CrossRefGoogle Scholar
  53. Tsujimoto Y, Shimizu S (2007) Role of the mitochondrial membrane permeability transition in cell death. Apoptosis 12:835–840CrossRefGoogle Scholar
  54. Wang L, Wei D, Huang S, Peng Z, Le X, Wu TT, Yao J, Ajani J, Xie K (2003) Transcription factor Sp1 expression is a significant predictor of survival in human gastric cancer. Clin Cancer Res 9:6371–6380Google Scholar
  55. Wang XB, Peng WQ, Yi ZJ, Zhu SL, Gan QH (2007) Expression and prognostic value of transcriptional factor sp1 in breast cancer. Ai Zheng 26:996–1000Google Scholar
  56. Wang X, Wang J, Lin S, Geng Y, Wang J, Jiang B (2008) Sp1 is involved in H2O2-induced PUMA gene expression and apoptosis in colorectal cancer cells. J Exp Clin Cancer Res 27:44CrossRefGoogle Scholar
  57. Yao JC, Wang L, Wei D, Gong W, Hassan M, Wu TT, Mansfield P, Ajani J, Xie K (2004) Association between expression of transcription factor Sp1 and increased vascular endothelial growth factor expression, advanced stage, and poor survival in patients with resected gastric cancer. Clin Cancer Res 10:4109–4117CrossRefGoogle Scholar
  58. Yao D, Cui H, Zhou S, Guo L (2017) Morin inhibited lung cancer cells viability, growth, and migration by suppressing miR-135b and inducing its target CCNG2. Tumour Biol 39:1010428317712443Google Scholar

Copyright information

© The Pharmaceutical Society of Korea 2019

Authors and Affiliations

  1. 1.Molecular Cancer ResearchSoonchunhyang University College of MedicineCheonanRepublic of Korea
  2. 2.Department of DermatologySoonchunhyang University HospitalSeoulRepublic of Korea

Personalised recommendations