5-Aza Cytidine Sensitizes HCT116 Colorectal Cancer Cells to 5-Fluorouracil: Enhanced Efficacy and Altered Genomic Variation

• Hussein Sabit

  Department of Medical Biotechnology, College of Biotechnology, Misr University for Science and Technology, P. O. Box 77, Giza, Egypt

• Thamer Alshammari

  Genetic Research Department, Institute for Research and Medical Consultations, Imam Abdulrahman Bin Faisal University, P. O. Box 1982, Dammam, 31441 Saudi Arabia

• Shimaa Abdel-Ghany

  Department of Environmental Biotechnology, College of Biotechnology, Misr University for Science and Technology, P. O. Box 77, Giza, Egypt

• Mohamed Berika

  Rehabilitation Science Department, College of Applied Medical Sciences, King Saud University, Riyadh, Saudi Arabia

• Hirendra Banerjee

  Elizabeth City State University Campus of the University of North Carolina, Elizabeth City, NC 27909, USA

• Ahmed El-Hashash

  Elizabeth City State University Campus of the University of North Carolina, Elizabeth City, NC 27909, USA

The escalating global burden of cancer, with a near 40% mortality increase over four decades and a projected 60% surge this decade, culminating in an estimated 13 million deaths by 2030, underscores the urgent need for improved therapies. Colorectal cancer (CRC), the third most frequent malignancy and second leading cause of cancer mortality worldwide, accounting for approximately 10% of diagnoses, necessitates innovative treatment strategies. This in vitro study investigated the effects of combined exposure to 5-fluorouracil (5-FU) and 5-Aza cytidine (5-Aza) on human colorectal carcinoma cells (HCT116). Employing MTT, wound healing, and colony formation assays, we observed significant reductions in cell viability, migration, and proliferation with the combined treatment compared to monotherapies. Whole-exome sequencing (WES) revealed a substantial decrease in the number and diversity of single-nucleotide polymorphisms (SNPs) in 5-FU-treated cells. These findings demonstrate that combined 5-Aza/5-FU exposure alters HCT116 cellular responses; however, formal pharmacological synergy cannot be inferred from the present experimental design. The synergistic effect of 5-Aza sensitized CRC cells to 5-FU.

[1] Kuipers EJ, Rösch T, Bretthauer M. Colorectal cancer screening--optimizing current strategies and new directions. Nat Rev Clin Oncol 2013; 10(3): 130-42.

[2] Bhandari A, Woodhouse M, Gupta S. Colorectal cancer is a leading cause of cancer incidence and mortality among adults younger than 50 years in the USA: a SEER-based analysis with comparison to other young-onset cancers. Journal of investigative medicine 2017; 65(2): 311-315.

[3] Sung H, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA: A Cancer Journal for Clinicians 2021; 71(3): 209-249.

[4] Kuipers EJ, et al. Colorectal cancer. Nature reviews. Disease primers 2015; 1: 15065-15065.

[5] AlDubayan SH, et al. Inherited DNA-Repair Defects in Colorectal Cancer. American journal of human genetics 2018; 102(3): 401-414.

[6] Huck MB, Bohl JL. Colonic Polyps: Diagnosis and Surveillance. Clinics in colon and rectal surgery 2016; 29(4): 296-305.

[7] Bonnington SN, Rutter MD. Surveillance of colonic polyps: Are we getting it right? World journal of gastroenterology 2016; 22(6): 1925-1934.

[8] Baharudin R, et al. Identification of Predictive DNA Methylation Biomarkers for Chemotherapy Response in Colorectal Cancer. Frontiers in Pharmacology 2017; 8.

[9] Vodenkova S, et al. 5-fluorouracil and other fluoropyrimidines in colorectal cancer: Past, present and future. Pharmacol Ther 2020; 206: 107447.

[10] Lee JJ, Beumer JH, Chu E. Therapeutic drug monitoring of 5-fluorouracil. Cancer Chemother Pharmacol 2016; 78(3): 447-64.

[11] Fakhr E, et al. LEF1 silencing sensitizes colorectal cancer cells to oxaliplatin, 5-FU, and irinotecan. Biomed Pharmacother 2021; 143: 112091.

[12] Zhang Q, et al. Exosomal transfer of p-STAT3 promotes acquired 5-FU resistance in colorectal cancer cells. J Exp Clin Cancer Res 2019; 38(1): 320.

[13] Sethy C, Kundu CN. 5-Fluorouracil (5-FU) resistance and the new strategy to enhance the sensitivity against cancer: Implication of DNA repair inhibition. Biomedicine & Pharmacotherapy 2021; 137: 111285.

[14] Mindt S, et al. Therapeutic drug monitoring (TDM) of 5-fluorouracil (5-FU): new preanalytic aspects. Clin Chem Lab Med 2019; 57(7): 1012-1016.

[15] Mossman D, Kim KT, Scott RJ. Demethylation by 5-aza-2'-deoxycytidine in colorectal cancer cells targets genomic DNA whilst promoter CpG island methylation persists. BMC Cancer 2010; 10: 366.

[16] Tsai HC, et al. Transient low doses of DNA-demethylating agents exert durable antitumor effects on hematological and epithelial tumor cells. Cancer Cell 2012; 21(3): 430-46.

[17] Ishiguro M, et al. Effect of combined therapy with low-dose 5-aza-2'-deoxycytidine and irinotecan on colon cancer cell line HCT-15. Ann Surg Oncol 2007; 14(5): 1752-62.

[18] Tsai HC, et al. Transient low doses of DNA-demethylating agents exert durable antitumor effects on hematological and epithelial tumor cells. Cancer Cell 2012; 21(3): 430-46.

[19] He J, et al. Chemoresistance of colorectal cancer to 5-fluoro-uracil is associated with silencing of the BNIP3 gene through aberrant methylation. J Cancer 2017; 8(7): 1187-1196.

[20] Morita S, et al. The Synergistic Effect of 5-Aza-2′-Deoxycytidine and 5-Fluorouracil on Drug-Resistant Tumors. Oncology 2006; 71(5-6): 437-445.

[21] Sanaei M, Kavoosi F, Ghasemzadeh V. Investigation of the Effect of 5-Aza-2'-Deoxycytidine in Comparison to and in Combination with Trichostatin A on p16INK4a, p14ARF, p15INK4b Gene Expression, Cell Growth Inhibition and Apoptosis Induction in Colon Cancer Caco-2 Cell Line. Int J Prev Med 2021; 12: 64.

[22] Huang YJ, et al. Antrodia cinnamomea Enhances Chemo-Sensitivity of 5-FU and Suppresses Colon Tumorigenesis and Cancer Stemness via Up-Regulation of Tumor Suppressor miR-142-3p. Biomolecules 2019; 9(8).

[23] Zhu Y, et al. Inhibition of CDK1 Reverses the Resistance of 5-Fu in Colorectal Cancer. Cancer Manag Res 2020; 12: 11271-11283.

[24] Gu XY, et al. Over-expression of EGFR regulated by RARA contributes to 5-FU resistance in colon cancer. Aging (Albany NY) 2020; 12(1): 156-177.

[25] Chen XL, et al. The effect of two nucleoside antitumor drugs on the proliferation and DNA methylation of human gastric cancer cells. Oncol Lett 2015; 10(3): 1919-1923.

[26] Chen XB, et al. [Effects of 5-Aza-dC on 5-Fu chemosensitivity by modulating TIP30 gene expression in human colorectal cancer cells]. Zhonghua Zhong Liu Za Zhi 2013; 35(11): 824-7.

[27] Ghafouri-Fard S, et al. 5-Fluorouracil: A Narrative Review on the Role of Regulatory Mechanisms in Driving Resistance to This Chemotherapeutic Agent. Front Oncol 2021; 11: 658636.

[28] Futscher BW. 5-aza-2′Deoxycytidine, in Encyclopedia of Cancer, M. Schwab, Editor. 2011, Springer Berlin Heidelberg: Berlin, Heidelberg. p. 325-328.

[29] Țigu AB, et al. The Synergistic Antitumor Effect of 5-Fluorouracil Combined with Allicin against Lung and Colorectal Carcinoma Cells. Molecules 2020; 25(8).

[30] Shakibaei M, et al. Curcumin potentiates antitumor activity of 5-fluorouracil in a 3D alginate tumor microenvironment of colorectal cancer. BMC Cancer 2015; 15: 250.

[31] Chang HW, et al. 5-azacytidine induces anoikis, inhibits mammosphere formation and reduces metalloproteinase 9 activity in MCF-7 human breast cancer cells. Molecules 2014; 19(3): 3149-59.

[32] Gu X, et al. Decitabine- and 5-azacytidine resistance emerges from adaptive responses of the pyrimidine metabolism network. Leukemia 2021; 35(4): 1023-1036.

[33] Zhang J, et al. DNA Methyltransferases in Cancer: Biology, Paradox, Aberrations, and Targeted Therapy. Cancers 2020; 12(8): 2123.

[34] Pagel KA, et al. When loss-of-function is loss of function: assessing mutational signatures and impact of loss-of-function genetic variants. Bioinformatics (Oxford, England) 2017; 33(14): i389-i398.

[35] Havens MA, Duelli DM, Hastings ML. Targeting RNA splicing for disease therapy. Wiley interdisciplinary reviews. RNA 2013; 4(3): 247-266.

• PDF

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.