Muscadine grape skin extract reverts snail-mediated epithelial mesenchymal transition via superoxide species in human prostate cancer cells

BMC Complementary and Alternative Medicine

Valerie A Odero-Marah¹  Niren Murthy⁴  Kousik Kundu⁴  Tamaro Hudson²  Rebecca S Arnold³  Basil Smith¹  Petrina Barnett¹  Liza J Burton¹ 

[1] Center for Cancer Research and Therapeutic Development, Department of Biological Sciences, Clark Atlanta University, 223 James P Brawley Dr SW, 30314 Atlanta, GA, USA;Department of Medicine, Howard University, Washington, DC 20060, USA;Department of Pathology, Emory University School of Medicine, Washington, DC 20060, USA;Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia

关键词: Prostate cancer;    Superoxide;    Reactive oxygen species;    EMT;    Snail;    Muscadine grape skin extract;   

Others  :  1220226
DOI  :  10.1186/1472-6882-14-97

 received in 2013-12-11, accepted in 2014-02-28,  发布年份 2014

【 摘 要 】

Background

Snail transcription factor can induce epithelial-mesenchymal transition (EMT), associated with decreased cell adhesion-associated molecules like E-cadherin, increased mesenchymal markers like vimentin, leading to increased motility, invasion and metastasis. Muscadine grape skin extract (MSKE) has been shown to inhibit prostate cancer cell growth and induce apoptosis without affecting normal prostate epithelial cells. We investigated novel molecular mechanisms by which Snail promotes EMT in prostate cancer cells via Reactive Oxygen Species (ROS) and whether it can be antagonized by MSKE.

Methods

ARCaP and LNCaP cells overexpressing Snail were utilized to examine levels of reactive oxygen species (ROS), specifically, superoxide, in vitro using Dihydroethidium (DHE) or HydroCy3 dyes. Mitosox staining was performed to determine whether the source of ROS was mitochondrial in origin. We also investigated the effect of Muscadine grape skin extract (MSKE) on EMT marker expression by western blot analysis. Migration and cell viability using MTS proliferation assay was performed following MSKE treatments.

Results

Snail overexpression in ARCaP and LNCaP cells was associated with increased concentration of mitochondrial superoxide, in vitro. Interestingly, MSKE decreased superoxide levels in ARCaP and LNCaP cells. Additionally, MSKE and Superoxide Dismutase (SOD) reverted EMT as evidenced by decreased vimentin levels and re-induction of E-cadherin expression in ARCaP-Snail cells after 3 days, concomitant with reduced cell migration. MSKE also decreased Stat-3 activity in ARCaP-Snail cells.

Conclusions

This study shows that superoxide species may play a role in Snail transcription factor-mediated EMT. Therefore, therapeutic targeting of Snail with various antioxidants such as MSKE may prove beneficial in abrogating EMT and ROS-mediated tumor progression in human prostate cancer.

【 授权许可】

   
2014 Burton et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Greenspan P, Bauer J, Stanley H, Pollock SH, Gangemi JD, Mayer EP, Ghaffar A, Hargrove JL, Hartle DK: Antiinflammatory properties of the muscadine grape (vitis rotundifolia). J Agric Food Chem 2005, 53(22):8481-8484.
• [2]He F, Mu L, Yan G, Liang N, Pan Q, Wang J, Reeves M, Duan C: Biosynthesis of anthocyanins and their regulation in colored grapes. Molecules 2010, 15:9057-9091.
• [3]Lee JHTS: Ellagic acid and ellagitannins affect on sedimentation in muscadine juice and wine. J Agric Food Chem 2002, 50:3971-3976.
• [4]Hudson TS, Hartle DK, Hursting SD, Nunez NP, Wang TT, Young HA, Arany P, Green JE: Inhibition of prostate cancer growth by muscadine grape skin extract and resveratrol through distinct mechanisms. Cancer Res 2007, 67(17):8396-8405.
• [5]Wang SYJH: Scavenging capacity of berry crops on superoxide radicals, hydrogen peroxide, hydroxyl radicals, and singlet oxygen. J Agric Food Chem 2000, 48:5677-5684.
• [6]Liou GY, Storz P: Reactive oxygen species in cancer. Free Radic Res 2010, 44(5):479-496.
• [7]Khandrika L, Kumar B, Koul S, Maroni P, Koul HK: Oxidative stress in prostate cancer. Cancer Lett 2009, 282(2):125-136.
• [8]Fleshner NE, Klotz LH: Diet, androgens, oxidative stress and prostate cancer susceptibility. Cancer Metastasis Rev 1998, 17(4):325-330.
• [9]Stolarek RA, Potargowicz E, Seklewska E, Jakubik J, Lewandowski M, Jeziorski A, Nowak D: Increased H2O2 level in exhaled breath condensate in primary breast cancer patients. J Cancer Res Clin Oncol 2010, 136(6):923-930.
• [10]Haklar G, Sayin-Ozveri E, Yuksel M, Aktan AO, Yalcin AS: Different kinds of reactive oxygen and nitrogen species were detected in colon and breast tumors. Cancer Lett 2001, 165(2):219-224.
• [11]Rhyu DY, Yang Y, Ha H, Lee GT, Song JS, Uh ST, Lee HB: Role of reactive oxygen species in TGF-beta1-induced mitogen-activated protein kinase activation and epithelial-mesenchymal transition in renal tubular epithelial cells. J Am Soc Nephrol 2005, 16(3):667-675.
• [12]Radisky DC, Levy DD, Littlepage LE, Liu H, Nelson CM, Fata JE, Leake D, Godden EL, Albertson DG, Nieto MA, Werb Z, Bissell MJ: Rac1b and reactive oxygen species mediate MMP-3-induced EMT and genomic instability. Nature 2005, 436(7047):123-127.
• [13]Barnett P, Arnold RS, Mezencev R, Chung LW, Zayzafoon M, Odero-Marah V: Snail-mediated regulation of reactive oxygen species in ARCaP human prostate cancer cells. Biochem Biophys Res Commun 2011, 404(1):34-39.
• [14]Cano A, Perez-Moreno MA, Rodrigo I, Locascio A, Blanco MJ, del Barrio MG, Portillo F, Nieto MA: The transcription factor snail controls epithelial-mesenchymal transitions by repressing E-cadherin expression. Nat Cell Biol 2000, 2(2):76-83.
• [15]Neal CL, McKeithen D, Odero-Marah VA: Snail negatively regulates cell adhesion to extracellular matrix and integrin expression via the MAPK pathway in prostate cancer cells. Cell Adh Migr 2011, 5(3):249-257.
• [16]Peiro S, Escriva M, Puig I, Barbera MJ, Dave N, Herranz N, Larriba MJ, Takkunen M, Franci C, Munoz A, Virtanen I, Baulida J, Garcia de Herreros A: Snail1transcriptional repressor binds to its own promoter and controls its expression. Nucleic Acids Res 2006, 34:2077-2084.
• [17]Smith BN, Odero-Marah VA: The role of snail in prostate cancer. Cell Adh Migr 2012, 6(5):433-41.
• [18]Vega S, Morales AV, Ocana OH, Valdés F, Fabregat I, Nieto MA: Snail blocks the cell cycle and confes resistance to cell death. Genes Dev 2004, 18:1131-1143.
• [19]Zhau H, Odero-Marah VA, Lue H, Nomura T, Wang R, Chu G, Liu ZR, Zhou BP, Huang WC, Chung LW: Epithelial to mesenchymal transition (EMT) in human prostate cancer: lessons learned from ARCaP model. Clin Exp Metastasis 2008, 25:601-610.
• [20]Turkson J, Jove R: STAT proteins: novel molecular targets for cancer drug discovery. Oncogene 2000, 19:6613-6626.
• [21]Odero-Marah VA, Wang R, Chu G, Zayzafoon M, Xu J, Shi C, Marshall FF, Zhau HE, Chung LW: Receptor activator of NF-kappaB Ligand (RANKL) expression is associated with epithelial to mesenchymal transition in human prostate cancer cells. Cell Res 2008, 18(8):858-870.
• [22]McKeithen D, Graham T, Chung LW, Odero-Marah V: Snail transcription factor regulates neuroendocrine differentiation in LNCaP prostate cancer cells. Prostate 2010, 70(9):982-992.
• [23]Xu J, Wang R, Xie ZH, Odero-Marah V, Pathak S, Multani A, Chung LW, Zhau HE: Prostate cancer metastasis: role of the host microenvironment in promoting epithelial to mesenchymal transition and increased bone and adrenal gland metastasis. Prostate 2006, 66(15):1664-1673.
• [24]Zhau HY, Chang SM, Chen BQ, Wang Y, Zhang H, Kao C, Sang QA, Pathak SJ, Chung LW: Androgen-repressed phenotype in human prostate cancer. Proc Natl Acad Sci U S A 1996, 93(26):15152-15157.
• [25]Xia C, Meng Q, Liu LZ, Rojanasakul Y, Wang XR, Jiang BH: Reactive oxygen species regulate angiogenesis and tumor growth through vascular endothelial growth factor. Cancer Res 2007, 67(22):10823-10830.
• [26]Simon AR, Rai U, Fanburg BL, Cochran BH: Activation of the JAK-STAT pathway by reactive oxygen species. Am J Physiol 1998, 275:C1640-C1652.
• [27]Lim SD, Sun C, Lambeth JD, Marshall F, Amin M, Chung L, Petros JA, Arnold RS: Increased Nox1 and hydrogen peroxide in prostate cancer. Prostate 2005, 62(2):200-207.
• [28]Boyer B, Valles AM, Edme N: Induction and regulation of epithelial-mesenchymal transitions. Biochem Pharmacol 2000, 60(8):1091-1099.
• [29]Dong R, Lu JG, Wang Q, He XL, Chu YK, Ma QJ: Stabilization of Snail by HuR in the process of hydrogen peroxide induced cell migration. Biochem Biophys Res Commun 2007, 356(1):318-321.
• [30]Yi W, Fisher J, Akoh C: Study of anticancer activities of muscadine grape phenolics in vitro. J Agric Food Chem 2005, 53(22):8804-8812.
• [31]Fan F, Samuel S, Evans KW, Lu J, Xia L, Zhou Y, Sceusi E, Tozzi F, Ye XC, Mani SA, Ellis LM: Overexpression of Snail induces epithelial-mesenchymal transition and a cancer stem cell like phenotype in human colorectal cancer cells. Cancer Med 2013, 1(1):6-16.