Synergistic Cytotoxicity of 5-Fluorouracil and Epigallocatechin-3-Gallate on Colorectal Cancer Stem Cell
Abstract
Background: Colorectal cancer stem cells (CR-CSCs) derived from the HCT-116 cell line established human colon carcinoma cell line, validated with CD44+/CD133+. The study investigates the synergistic effects of combining epigallocatechin gallate (EGCG) with 5-fluorouracil (5-FU) on CR-CSCs through comprehensive cytotoxicity assessments, aiming to enhance therapeutic outcomes. EGCG is a polyphenol with anti-cancer activity in green tea. Previous studies have reported that the anti-cancer activity of EGCG involves inhibition of proliferation and induction of apoptosis thereby reducing recurrence by as much as 51.6% in patients with colorectal adenoma after polypectomy. The significance lies in optimizing treatment strategies by understanding the potential synergies between conventional chemotherapeutic agents and natural compounds. Given 5-FU's status as a cornerstone in CR-CSCs chemotherapy and EGCG's emergence as a promising natural compound, the study delves into their individual and combined cytotoxicity profiles.
Methods: The single and combination assay aimed to determine the cytotoxicity of EGCG and 5-FU, including establishing the half inhibitory concentration (IC50) and combination index (CI) values. CR-CSCs colonies were disassociated, counted, and cultured in 96-well plates. Test solutions of varying concentrations were applied, and subsequent steps involved incubation, media removal, washing, MTT reagent addition, and absorbance measurement.
Results: The single cytotoxicity tests established individual IC50 values, revealing 141.26 µM for 5-FU and 464.56 µM for EGCG. Subsequent combination cytotoxicity tests demonstrated a synergistic effect at specific doses, indicated by CI values below 1.
Conclusions: These findings highlight the potential for increased cytotoxicity against CR-CSCs when treated with the combination of 5-FU and EGCG.
Keywords
DOI: 10.33371/ijoc.v18i2.1211
Article Metrics
Abstract View: 190, 
PDF Download: 149
References
Rawla P, Sunkara T, Barsouk A. Epidemiology of colorectal cancer: incidence, mortality, survival, and risk factors. Prz Gastroenterol. 2019;14(2):89-103.
Rubesin SE. Colon: Globocan 2020. Pract Fluoroscopy GI GU Tracts. 2020;419:169–90.
Bian J, Dannappel M, Wan C, Firestein R. Transcriptional Regulation of Wnt/β-Catenin Pathway in Colorectal Cancer. Cells. 2020;9(9):1–29.
Vogel JD, Eskicioglu C, Weiser MR, et al. The American society of colon and rectal surgeons clinical practice guidelines for the treatment of colon cancer. Dis Colon Rectum. 2019;60(10):999–1017.
Zhu J, Wang S, Chen Y, et al. miR-19 targeting of GSK3β mediates sulforaphane suppression of lung cancer stem cells. J Nutr Biochem. 2017 Jun;44:80-91.
Vaillancourt K, Ben Lagha A, Grenier D. A green tea extract and epigallocatechin-3-gallate attenuate the deleterious effects of irinotecan in an oral epithelial cell model. Arch Oral Biol. 2021 Jun 1;126.
Cho YH, Ro EJ, Yoon JS, et al. 5-FU promotes stemness of colorectal cancer via p53-mediated WNT/β-catenin pathway activation. Nat Commun. 2020 Oct 21;11(1):5321.
Blondy S, David V, Verdier M, et al. 5-Fluorouracil resistance mechanisms in colorectal cancer: From classical pathways to promising processes. Cancer Sci. 2020 Sep;111(9):3142-3154.
Sethy C, Kundu CN. 5-Fluorouracil [5-FU] resistance and the new strategy to enhance the sensitivity against cancer: Implication of DNA repair inhibition. Biomed Pharmacother. 2021 May 1;137:111285.
Wang J, Wang W, Cai H, et al. MACC1 facilitates chemoresistance and cancer stem cell-like properties of colon cancer cells through the PI3K/AKT signaling pathway. Mol Med Rep. 2019;16(6):8747–54.
Shin YS, Kang SU, Park JK, et al. Anti-cancer effect of [-]-epigallocatechin-3-gallate [EGCG] in head and neck cancer through repression of transactivation and enhanced degradation of β-catenin. Phytomedicine. 2018 Nov 15;23(12):1344–55.
Hu F, Wei F, Wang Y, et al. EGCG synergizes the therapeutic effect of cisplatin and oxaliplatin through autophagic pathway in human colorectal cancer cells. J Pharmacol Sci. 2019 May 1;128(1):27–34.
Thakur VS, Ruhul Amin ARM, Paul RK, et al. p53-Dependent p21-mediated growth arrest pre-empts and protects HCT116 cells from PUMA-mediated apoptosis induced by EGCG. Cancer Lett. 2017 Oct 28;296(2):225–32.
Kwon KR, Alam MB, Park JH, et al. Attenuation of UVB-induced photo-aging by polyphenolic-rich spatholobus suberectus stem extract via modulation of MAPK/AP-1/MMPs signaling in human keratinocytes. Nutrients. 2019;11(6).
Ma YS, Li W, Liu Y, et al. Targeting colorectal cancer stem cells as an effective treatment for colorectal cancer. Technol Cancer Res Treat. 2020;19(1):1–9.
Rey I, Putra A, Lindarto D, Yusuf F. Association between CD133 expression and clinicopathological profile in colorectal cancer. Med Glas. 2020;17(2):304–9.
Rabinovich I, Sebastião APM, Lima RS, et al. Cancer stem cell markers ALDH1 and CD44+/CD24– phenotype and their prognosis impact in invasive ductal carcinoma. Eur J Histochem. 2018;62(3):231–7.
Gemei M, Di Noto R, Mirabelli P, Del Vecchio L. Cytometric profiling of CD133+ cells in human colon carcinoma cell lines identifies a common core phenotype and cell type-specific mosaics. Int J Biol Markers. 2018;28(3):267–73.
Morata-Tarifa C, Jiménez G, García MA, et al. Low adherent cancer cell subpopulations are enriched in tumorigenic and metastatic epithelial-to-mesenchymal transition-induced cancer stem-like cells. Sci Rep. 2017;6(May 2017):1–13.
Yu J, He Z, He X, et al. Comprehensive Analysis of the Expression and Prognosis for MMPs in Human Colorectal Cancer. Front Oncol. 2021;11(November):1–13.
Hermansyah D, Putra A, Munir D, et al. Synergistic Effect of Curcuma longa Extract in Combination with Phyllanthus niruri Extract in Regulating Annexin A2, Epidermal Growth Factor Receptor, Matrix Metalloproteinases, and Pyruvate Kinase M1/2 Signaling Pathway on Breast Cancer Stem Cell. Open Access Maced J Med Sci. 2021 May 9;9[A]:271–85.
Luo KW, Xia J, Cheng BH, et al. Tea polyphenol EGCG inhibited colorectal-cancer-cell proliferation and migration via downregulation of STAT3. Gastroenterol Rep. 2021;9(1):59–70.
La X, Zhang L, Li Z, et al. (-)-Epigallocatechin Gallate (EGCG) Enhances the Sensitivity of Colorectal Cancer Cells to 5-FU by Inhibiting GRP78/NF-κB/miR-155-5p/MDR1 Pathway. J Agric Food Chem. 2019 Mar 6;67(9):2510-2518.
Nurrachma MY, Maran GG, Putri NB, et al. Fingerroot [Boesenbergia pandurata] Extract Inhibits Proliferation and Migration of 4T1 Metastatic Breast Cancer Cells. Indones J Cancer Chemoprevention. 2020;11(3):103-14.
Li XX, Liu C, Dong SL, et al. Anticarcinogenic potentials of tea catechins. Front Nutr. 2022 Dec 5;9:1060783.
Kumazaki M, Noguchi S, Yasui Y, et al. Anti-cancer effects of naturally occurring compounds through modulation of signal transduction and miRNA expression in human colon cancer cells. J Nutr Biochem. 2013 Nov;24(11):1849-58.
Caglar HO, Biray Avci C. Alterations of cell cycle genes in cancer: unmasking the role of cancer stem cells. Mol Biol Rep. 2020;47(4):3065–76.
Zhou Y, Xia L, Wang H, et al. Cancer stem cells in progression of colorectal cancer. Oncotarget. 2019;9(70):33403–15.
Garza-Treviño EN, Said-Fernández SL, Martínez-Rodríguez HG. Understanding the colon cancer stem cells and perspectives on treatment. Cancer Cell Int. 2017;15[1]:1–9.
Bai X, Ni J, Beretov J, et al. Cancer stem cell in breast cancer therapeutic resistance. Cancer Treat Rev. 2018 Sep;69:152-163.
Refbacks
- There are currently no refbacks.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Email this article 
