Gallic Acid, a Promising Chemopreventive Agent, Antagonizes Breast Cancer: Molecular Mechanisms of Action from Cell Culture Studies

Hanaa H Ahmed

Abstract

Objective The present consideration converged on trailing the underlying molecular mechanism of gallic acid in striking the distinctly invasive breast cancer cell line; MDA-MB-231 in vitro. Methods The cytotoxic influence of gallic acid and 5-fluorouracil (5-FU) on MDA-MB-231 cells was stipulated by MTT assay. The expression levels of apoptosis, proliferation, progression, invasion and metastases-related genes were identified by RT-PCR. Results Our data specified that gallic acid and 5-FU displayed modest growth suppression with IC50 values of 251 and 367.171 µM contra MDA-MB-231 cells respectively after 48 h. Otherwise, treatment of MDA-MB-231 cells with gallic acid inspired significant downregulation in the expression level of survivin, STAT3, IL-6, VEGF, Slug, Snail, MMP7 and Notch-4 genes. Whereas, 5-FU instigated significant downregulation in the expression level survivin, STAT3, IL-6 and VEGF genes correlated with significant upregulation in the expression level of Snail and Notch-4 genes. Likewise, 5-FU provoked insignificant upregulation in the expression level of Slug and downregulation in the expression level MMP7 genes. Conclusion The attained data throw light on the distinct mechanism by which gallic acid could engage breast cancer. Gallic acid manifested chemopreventive potentiality against breast cancer aggressiveness through rectifying several dysregulated molecules implicated in cancer cell survival, proliferation, progression, angiogenesis, chemoresistance and metastasis.

Full Text:

PDF

References

Cao L, Yang Y, Ye Z, Lin B, Zeng J, Li C, et al. Quercetin‑3‑methyl ether suppresses human breast cancer stem cell formation by inhibiting the Notch1 and PI3K/Akt signalling pathways. Int J Mol Med 2018;24:1625–1636.

Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics, 2009. CA Cancer J Clin 2009;59:225–249.

Felipe Lima J, Nofech-Mozes S, Bayani J, Bartlett J. EMT in Breast Carcinoma—A Review. J Clin Med 2016;5:65.

Asiedu MK, Beauchamp-Perez FD, Ingle JN, Behrens MD, Radisky DC, Knutson KL. AXL induces epithelial-to-mesenchymal transition and regulates the function of breast cancer stem cells. Oncogene 2014;33:1316–1324.

Czerwinska P, Kaminska B. Review Regulation of breast cancer stem cell features. Contemp Oncol 2015;19:A7-A15.

Holohan C, Van Schaeybroeck S, Longley DB, Johnston PG. Cancer drug resistance: an evolving paradigm. Nat Rev Cancer 2013;13:714–726.

Li X, Kong X, Kong X, Wang Y, Yan S, Yang Q. 53BP1 Sensitizes breast cancer cells to 5-Fluorouracil. PLoS ONE 2013;8(9):e74928.

Longley DP, Harkin DP, Johnston PG. 5-Fluorouracil: Mechanisms of action and clinical strategies. Nature Reviews 2003;3:330-338.

Kufe DW, Major PP. 5-Fluorouracil incorporation into human breast carcinoma RNA correlates with cytotoxicity. J Biol Chem1981;256:9802–9805.

Afzal M, Safer AM, Menon M. Green tea polyphenols and their potential role in health and disease. Inflammopharmacology 2015;23:151–161.

Sourani Z, Pourgheysari B, Beshkar P, Shirzad H, Shirzad M. Gallic Acid Inhibits Proliferation and Induces Apoptosis in Lymphoblastic Leukemia Cell Line (C121). Iran J Med Sci 2016;41:525-530.

Chen H-M, Wu Y-C, Chia Y-C, Chang F-R, Hsu H-K, Hsieh Y-C, et al. Gallic acid, a major component of Toona sinensis leaf extracts, contains a ROS-mediated anti-cancer activity in human prostate cancer cells. Cancer Lett 2009;286:161–171.

Ohno T, Inoue M, Ogihara Y. Cytotoxic activity of gallic acid against liver metastasis of mastocytoma cells P-815. Anticancer Res 2001;21:3875–3880.

Maggi-Capeyron M-F, Ceballos P, Cristol J-P, Delbosc S, Le Doucen C, Pons M, et al. Wine Phenolic Antioxidants Inhibit AP-1 Transcriptional Activity. J Agric Food Chem 2001;49:5646–5652.

Liu Z, Schwimer J, Liu D, Lewis J, Greenway FL, York DA, et al. Gallic acid is partially responsible for the antiangiogenic activities of Rubus leaf extract. Phytother Res 2006;20:806–813.

Nowakowska A, Tarasiuk J. Comparative effects of selected plant polyphenols, gallic acid and epigallocatechin gallate, on matrix metalloproteinases activity in multidrug resistant MCF7/DOX breast cancer cells. Acta Biochim Pol 2016;63:571–575.

Pandya K, Meeke K, Clementz AG, Rogowski A, Roberts J, Miele L, et al. Targeting both Notch and ErbB-2 signalling pathways is required for prevention of ErbB-2-positive breast tumor recurrence. Br J Cancer 2011;105:796–806.

Van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: The MTT assay. Methods Mol Biol 2011;731:237-245.

Speth PA, Linssen PC, Boezeman JB, Wessels HM, Haanen C. Cellular and plasma adriamycin concentrations in long-term infusion therapy of leukemia patients. Cancer Chemother Pharmacol 1987;20:305–310.

Gyamfi J, Lee Y-H, Eom M, Choi J. Interleukin-6/STAT3 signalling regulates adipocyte induced epithelial-mesenchymal transition in breast cancer cells. Sci Rep 2018;8:8859.

Lee K, Nam M, Lee H, Hong C, Lee K. Protective effects of chebulic acid on alveolar epithelial damage induced by urban particulate matter. BMC Complement Altern Med 2017;17:373.

Liu L-Z, Jiang Y, Carpenter RL, Jing Y, Peiper SC, Jiang B-H. Role and Mechanism of Arsenic in Regulating Angiogenesis. PLoS ONE 2011;6:e20858.

Han Y-H, Kee J-Y, Hong S-H. Rosmarinic Acid Activates AMPK to Inhibit Metastasis of Colorectal Cancer. Front Pharmacol 2018;9:68.

Gong C, Shen J, Fang Z, Qiao L, Feng R, Lin X, et al. Abnormally expressed JunB transactivated by IL-6/STAT3 signaling promotes uveal melanoma aggressiveness via epithelial-mesenchymal transition. Biosci Rep 2018;38:BSR20180532.

Di D, Chen L, Guo Y, Wang L, Zhao C, Ju J. BCSC-1 suppresses human breast cancer metastasis by inhibiting NF-κB signaling. Int J Oncol 2018;52:1674-1684.

Upadhyay P, Sudhir N, Kaur E, Aich J, Dani P, Sethunath V, et al. Notch pathway activation is essential for maintenance of stem-like cells in early tongue cancer. Oncotarget 2016;7:50437–50449.

Wang M-C, Wu A-G, Huang Y-Z, Shao G-L, Ji S-F, Wang R-W, et al. Autophagic regulation of cell growth by altered expression of Beclin 1 in triple-negative breast cancer. Int J Clin Exp Med 2015;8:7049–7058.

Kim H, Lee S, Park J-H, Lee S, Lee B-W, Hee Ihm S, et al. Enhanced protection of Ins-1 β cells from apoptosis under hypoxia by delivery of DNA encoding secretion signal peptide-linked exendin-4. J Drug Target 2009;17:242–248.

Li Q-X, Zhao J, Liu J-Y, Jia L-T, Huang H-Y, Xu Y-M, et al. Survivin stable knockdown by siRNA inhibits tumor cell growth and angiogenesis in breast and cervical cancers. Cancer Biol Ther 2006;5:860–866.

Li T, Chen G, Su C, Lin J, Yeh C, Cheng K, et al. Ellagic acid induced p53/p21 expression, G1 arrest and apoptosis in human bladder cancer T24 cells. Anticancer Res 2005;25:971–979.

Narayanan BA, Geoffroy O, Willingham MC, Re GG, Nixon DW. p53/p21(WAF1/CIP1) expression and its possible role in G1 arrest and apoptosis in ellagic acid treated cancer cells. Cancer Lett 1999;136:215–221.

Chen H-S, Bai M-H, Zhang T, Li G-D, Liu M. Ellagic acid induces cell cycle arrest and apoptosis through TGF-β/Smad3 signalling pathway in human breast cancer MCF-7 cells. Int J Oncol 2015;46:1730–1738.

Mobahat M, Narendran A, Riabowol K. Survivin as a Preferential Target for Cancer Therapy. Int J Mol Sci 2014;15:2494–2516.

Lee B-J, Chon K-M, Kim Y-S, An WG, Roh H-J, Goh E-K, et al. Effects of Cisplatin, 5-Fluorouracil, and Radiation on Cell Cycle Regulation and Apoptosis in the Hypopharyngeal Carcinoma Cell Line. Chemotherapy 2005;51:103–110.

Yang B, Eshleman JR, Berger NA, Markowitz SD. Wild-type p53 protein potentiates cytotoxicity of therapeutic agents in human colon cancer cells. Clin Cancer Res 1996;2:1649–1657.

Liu HC, Chen GG, Vlantis AC, Leung BCS, Tong MCF, van Hasselt CA. 5-fluorouracil mediates apoptosis and G1/S arrest in laryngeal squamous cell carcinoma via a p53-independent pathway. Cancer J 2006;12:482–493.

Kensara OA, El-Shemi AG, Mohamed AM, Refaat B, Idris S, Ahmad J. Thymoquinone subdues tumor growth and potentiates the chemopreventive effect of 5-fluorouracil on the early stages of colorectal carcinogenesis in rats. Drug Des Devel Ther 2016;10:2239–2253.

Cui X, Shen D, Kong C, Zhang Z, Zeng Y, Lin X, et al. NF-κB suppresses apoptosis and promotes bladder cancer cell proliferation by upregulating survivin expression in vitro and in vivo. Sci Rep 2017;7:40723.

Wang K, Brems JJ, Gamelli RL, Holterman AX. Survivin signaling is regulated through nuclear factor-kappa B pathway during glycochenodeoxycholate-induced hepatocyte apoptosis. Biochim Biophys Acta 2010;1803:1368–1375.

Heidarian E, Keloushadi M, Ghatreh-Samani K, Valipour P. The reduction of IL-6 gene expression, pAKT, pERK1/2, pSTAT3 signalling pathways and invasion activity by gallic acid in prostate cancer PC3 cells. Biomed Pharmacother 2016;84:264–269.

Neurath MF, Finotto S. IL-6 signalling in autoimmunity, chronic inflammation and inflammation-associated cancer. Cytokine Growth Factor Rev 2011;22:83–89.

Yu H, Pardoll D, Jove R. STATs in cancer inflammation and immunity: a leading role for STAT3. Nat Rev Cancer 2009;9:798–809.

Umesalma S, Sudhandiran G. Differential Inhibitory Effects of the Polyphenol Ellagic Acid on Inflammatory Mediators NF-κB, iNOS, COX-2, TNF-α, and IL-6 in 1,2-Dimethylhydrazine-Induced Rat Colon Carcinogenesis. Basic Clin Pharmacol Toxicol 2010;107:650–655.

Pandey A, Vishnoi K, Mahata S, Tripathi SC, Misra SP, Misra V, et al. Berberine and Curcumin Target Survivin and STAT3 in Gastric Cancer Cells and Synergize Actions of Standard Chemotherapeutic 5-Fluorouracil. Nutr Cancer 2015;67:1293–1304.

Hirano T, Ishihara K, Hibi M. Roles of STAT3 in mediating the cell growth, differentiation and survival signals relayed through the IL-6 family of cytokine receptors. Oncogene 2000;19:2548–2556.

Chung SS, Dutta P, Austin D, Wang P, Awad A, Vadgama JV. Combination of resveratrol and 5-flurouracil enhanced anti-telomerase activity and apoptosis by inhibiting STAT3 and Akt signalling pathways in human colorectal cancer cells. Oncotarget 2018;9:32943–32957.

Maida Y, Masutomi K. Telomerase reverse transcriptase moonlights: Therapeutic targets beyond telomerase. Cancer Sci 2015;106:1486–1492.

Bollrath J, Phesse TJ, von Burstin VA, Putoczki T, Bennecke M, Bateman T, et al. gp130-Mediated Stat3 Activation in Enterocytes Regulates Cell Survival and Cell-Cycle Progression during Colitis-Associated Tumorigenesis. Cancer Cell 2009;15:91–102.

Yin W, Cheepala S, Roberts JN, Syson-Chan K, DiGiovanni J, Clifford JL. Active Stat3 is required for survival of human squamous cell carcinoma cells in serum-free conditions. Mol Cancer 2006;5:15.

Selvendiran K, Bratasz A, Kuppusamy ML, Tazi MF, Rivera BK, Kuppusamy P. Hypoxia induces chemoresistance in ovarian cancer cells by activation of signal transducer and activator of transcription 3. Int J Cancer 2009;125:2198–2204.

Nguyen DP, Li J, Tewari AK. Inflammation and prostate cancer: the role of interleukin 6 (IL-6). BJU Int 2014;113:986–992.

Edderkaoui M, Odinokova I, Ohno I, Gukovsky I, Go VLW, Pandol SJ, et al. Ellagic acid induces apoptosis through inhibition of nuclear factor kappa B in pancreatic cancer cells. World J Gastroenterol 2008;14:3672–3680.

Yamamoto Y, Gaynor RB. Therapeutic potential of inhibition of the NF-κB pathway in the treatment of inflammation and cancer. J Clin Invest 2001;107:135–142.

Lee S-J, Lim K-T. Chemopreventive effect of plant originated glycoprotein on colitis-mediated colorectal cancer in A/J mice. J Biomed Sci 2007;15:111–121.

Azzolina A, Bongiovanni A, Lampiasi N. Substance P induces TNF-α and IL-6 production through NFκB in peritoneal mast cells. Biochim Biophys Acta 2003;1643:75–83.

Shibata Y, Baba E, Ariyama H, Arita S, Isobe T, Kusaba H, et al. Irinotecan-based combination chemotherapy for metastatic small intestinal adenocarcinoma. Oncol Lett 2010;1:423–426.

Li J, Cheng Y, Tai D, Martinka M, Welch DR, Li G. Prognostic significance of BRMS1 expression in human melanoma and its role in tumor angiogenesis. Oncogene 2011;30:896–906.

Aota K, Azuma M, Yamashita T, Tamatani T, Motegi K, Ishimaru N, et al. 5-Fluorouracil Induces Apoptosis through the Suppression of NF-κB Activity in Human Salivary Gland Cancer Cells. Biochem Biophys Res Commun 2000;273:1168–1174.

Yamamoto Y, Yin MJ, Lin KM, Gaynor RB. Sulindac inhibits activation of the NF-kappaB pathway. J Biol Chem 1999;274:27307–27314.

Karin M. NF-κB as a Critical Link Between Inflammation and Cancer. Cold Spring Harb Perspect Biol 2009;1:a000141.

Eichten A, Su J, Adler AP, Zhang L, Ioffe E, Parveen AA, et al. Resistance to Anti-VEGF Therapy Mediated by Autocrine IL6/STAT3 Signalling and Overcome by IL6 Blockade. Cancer Res 2016;76:2327–2339.

Kawano M, Hirano T, Matsuda T, Taga T, Horii Y, Iwato K, et al. Autocrine generation and requirement of BSF-2/IL-6 for human multiple myelomas. Nature 1988;332:83–85.

Gao SP, Mark KG, Leslie K, Pao W, Motoi N, Gerald WL, et al. Mutations in the EGFR kinase domain mediate STAT3 activation via IL-6 production in human lung adenocarcinomas. J Clin Invest 2007;117:3846–3856.

Hartman ZC, Poage GM, den Hollander P, Tsimelzon A, Hill J, Panupinthu N, et al. Growth of Triple-Negative Breast Cancer Cells Relies upon Coordinate Autocrine Expression of the Proinflammatory Cytokines IL-6 and IL-8. Cancer Res 2013;73:3470–3480.

Vanella L, Di Giacomo C, Acquaviva R, Barbagallo I, Li Volti G, Cardile V, et al. Effects of Ellagic Acid on Angiogenic Factors in Prostate Cancer Cells. Cancers 2013;5:726–738.

Bhat TA, Singh RB. Tumor angiogenesis – A potential target in cancer chemoprevention. Food Chem Toxicol 2008;46:1334–1345.

Kurebayashi J, Otsuki T, Kunisue H, Mikami Y, Tanaka K, Yamamoto S, et al. Expression of Vascular Endothelial Growth Factor (VEGF) Family Members in Breast Cancer. Jpn J Cancer Res 1999;90:977–981.

Wang N, Wang Z-Y, Mo S-L, Loo TY, Wang D-M, Luo H-B,et al. Ellagic acid, a phenolic compound, exerts anti-angiogenesis effects via VEGFR-2 signalling pathway in breast cancer. Breast Cancer Res Treat 2012;134:943–955.

Gingras D, Lamy S, Béliveau R. Tyrosine phosphorylation of the vascular endothelial-growth-factor receptor-2 (VEGFR-2) is modulated by Rho proteins. Biochem J 2000;348:273–280.

Bergers G, Brekken R, McMahon G, Vu TH, Itoh T, Tamaki K, et al. Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol 2000;2:737–744.

Ranger GS. Current concepts in colorectal cancer prevention with cyclooxygenase inhibitors. Anticancer Res 2014;34:6277–6282.

Niu G, Wright KL, Huang M, Song L, Haura E, Turkson J, et al. Constitutive Stat3 activity up-regulates VEGF expression and tumor angiogenesis. Oncogene 2002;21:2000–2008.

Anglesio MS, George J, Kulbe H, Friedlander M, Rischin D, Lemech C, et al. IL6-STAT3-HIF Signalling and Therapeutic Response to the Angiogenesis Inhibitor Sunitinib in Ovarian Clear Cell Cancer. Clin Cancer Res 2011;17:2538–2548.

Takahashi A, Watanabe T, Mondal A, Suzuki K, Kurusu-Kanno M, Li Z, et al. Mechanism-based inhibition of cancer metastasis with (−)-epigallocatechin gallate. Biochem Biophys Res Commun 2014;443:1–6.

Menon S, Beningo KA. Cancer Cell Invasion Is Enhanced by Applied Mechanical Stimulation. PLoS ONE 2011;6:e17277.

Gilles C, Polette M, Zahm JM, Tournier JM, Volders L, Foidart JM, et al. Vimentin contributes to human mammary epithelial cell migration. J Cell Sci 1999;112:4615–4625.

Inoue H, Takahashi H, Hashimura M, Eshima K, Akiya M, Matsumoto T, et al. Cooperation of Sox4 with β-catenin/p300 complex in transcriptional regulation of the Slug gene during divergent sarcomatous differentiation in uterine carcinosarcoma. BMC Cancer 2016;16:53.

Ng SH. Characterization of Colon Cancer Cell Culture Based Screening Assay to Study Effects of Phenolic Acids. Doctoral Dissertation, University of Saskatchewan. 2011; http://hdl.handle.net/10388/ETD-2011-09-163.

Chen Y, Zhou Z, Mo Q, Zhou G, Wang Y. Gallic Acid Attenuates Dimethylnitrosamine-Induced Liver Fibrosis by Alteration of Smad Phosphoisoform Signalling in Rats. BioMed Res Int 2018;2018:1–14.

Chaturvedi AK. Epidemiology and clinical aspects of HPV in head and neck cancers. Head Neck Pathol 2012;6:S16-24.

Umbreit C, Aderhold C, Faber A, Sommer JU, Sauter A, Hofheinz R, et al. Unexpected alteration of β-catenin and c-KIT expression by 5-FU and docetaxel in p16-positive squamous cell carcinoma compared to HPV-negative HNSCC cells in vitro. Anticancer Res 2013;33:2457–2465.

Huang W, Mao S, Zhang L, Lu B, Zheng L, Zhou F, et al. Phenolic compounds, antioxidant potential and antiproliferative potential of 10 common edible flowers from China assessed using a simulated in vitro digestion-dialysis process combined with cellular assays. J Sci Food Agric 2017;97:4760–4769.

Wang D, Chen Q, Tan Y, Liu B, Liu C. Ellagic acid inhibits human glioblastoma growth in vitro and in vivo. Oncol Rep 2017;37:1084–1092.

Ye X, Tam WL, Shibue T, Kaygusuz Y, Reinhardt F, Ng Eaton E, et al. Distinct EMT programs control normal mammary stem cells and tumor-initiating cells. Nature 2015;525:256–260.

Imani S, Hosseinifard H, Cheng J, Wei C, Fu J. Prognostic Value of EMT-inducing Transcription Factors (EMT-TFs) in Metastatic Breast Cancer: A Systematic Review and Meta-analysis. Scientific. Rep 2016;6:28587.

Romano G, Santi L, Bianco MR, Giuffrè MR, Pettinato M, Bugarin C, et al. The TGF-β pathway is activated by 5-fluorouracil treatment in drug resistant colorectal carcinoma cells. Oncotarget 2016;7:22077-22091.

Baruah MM, Khandwekar AP, Sharma N. Quercetin modulates Wnt signalling components in prostate cancer cell line by inhibiting cell viability, migration, and metastases. Tumor Biol 2016;37:14025–14034.

Palena C, Hamilton DH, Fernando RI. Influence of IL-8 on the epithelial–mesenchymal transition and the tumor microenvironment. Future Oncol 2012;8:713–722.

Chen Y-J, Chang L-S. Gallic acid downregulates matrix metalloproteinase-2 (MMP-2) and MMP-9 in human leukemia cells with expressed Bcr/Abl. Mol Nutr Food Res 2012;56:1398–1412.

Kim M, Murakami A, Ohigashi H. Modifying effects of dietary factors on (-)-epigallocatechin-3-gallate-induced pro-matrix metalloproteinase-7 production in HT-29 human colorectal cancer cells. Biosci Biotechnol Biochem 2007;71:2442–2450.

Zheng X, Jiang F, Katakowski M, Zhang X, Jiang H, Zhang ZG, et al. Sensitization of cerebral tissue in nude mice with photodynamic therapy induces ADAM17/TACE and promotes glioma cell invasion. Cancer Lett 2008;265:177–187.

Tsatas D, Kanagasundaram V, Kaye A, Novak U. EGF receptor modifies cellular responses to hyaluronan in glioblastoma cell lines. J Clin Neurosci 2002;9:282–288.

Fang Y, Pan Z, Li L, Lu Z, Zhang L, Wan D. MMP7 expression regulated by endocrine therapy in ERβ-positive colon cancer cells. J Exp Clin Cancer Res 2009;28:132.

Crawford HC, Fingleton BM, Rudolph-Owen LA, Goss KJ, Rubinfeld B, Polakis P, et al. The metalloproteinase matrilysin is a target of beta-catenin transactivation in intestinal tumors. Oncogene 1999;18: 2883–2891.

Liu Y, Tang Z, Lin Y, Qu X, Lv W, Wang G, et al. Effects of quercetin on proliferation and migration of human glioblastoma U251 cells. Biomed Pharmacother 2017;92:33–38.

Koch U, Radtke F. Notch and cancer: a double-edged sword. Cell Mol Life Sci 2007;64:2746–2762.

Wang D, Chen Q, Liu B, Li Y, Tan Y, Yang B. Ellagic acid inhibits proliferation and induces apoptosis in human glioblastoma cells. Acta Cir Bras 2016;31:143–149.

Wang Z, Li Y, Ahmad A, Azmi AS, Banerjee S, Kong D, et al. Targeting Notch signalling pathway to overcome drug resistance for cancer therapy. Biochim Biophys Acta 2010;1806:258–267.

Leong KG, Karsan A. Recent insights into the role of Notch signalling in tumorigenesis. Blood 2006;107: 2223–2233.

Li L-Q, Pan D, Zhang S-W, Xie Y-D, Zheng X-L, Chen H. Autophagy regulates chemoresistance of gastric cancer stem cells via the Notch signalling pathway. Eur Rev Med Pharmacol Sci 2018;22:3402–3407.

Refbacks

  • There are currently no refbacks.