In vitro anti-proliferative effect of capecitabine (Xeloda) combined with mocetinostat (MGCD0103) in 4T1 breast cancer cell line by immunoblotting

Document Type : Original Article

Authors

1 Department of Molecular Biology and Genetics, Faculty of Science and Letters, Bilecik Seyh Edebali University, Bilecik, Turkey

2 Biotechnology Research and Application Center, Bilecik Seyh Edebali University, Bilecik, Turkey

Abstract

Objective(s): Mouse breast cancer cell line 4T1 can accurately mimic the response to immune receptors and targeting therapeutic agents. Combined therapy has emerged as an important strategy with reduced side effects and maximum therapeutic effect. Mocetinostat (MGCD0103) is one of the members of Class I Histone Deacetylase Inhibitors (HDACi) and its mechanism of action has not been defined, yet. Capecitabine (Xeloda) is an antimetabolite and currently is widely utilized to treat a wide range of solid tumors. The aim of this study was to investigate the effects of the capecitabine, mocetinostat and their combined application on the 4T1 cell line.  
Materials and Methods: The effects of combined administration of mocetinostat and capecitabine on 4T1 cells were investigated by cell viability and migration assays, apoptosis analysis, and Western blotting technique.
Results: The concentrations of drugs that give a half-maximal response (IC50) were detected for capecitabine (1700 µM), mocetinostat (3,125 µM), and 50 µM Capecitabine+1,5 µM Mocetinostat for 48 hr. In capecitabine+mocetinostat combine group, we observed that cell migration decreased, DNA fragmentation increased compared to the control group. capecitabine + mocetinostat group induced apoptosis by decreasing Bcl-2, PI3K, Akt, c-myc protein levels, while increasing Bax, Caspase-3, PTEN, cleaved-PARP, Caspase-7, Caspase-9, p53, cleaved-Cas-9 protein levels in 4T1 cells. 
Conclusion: Capecitabine and mocetinostat played a toxic role through inducing apoptosis on 4T1 cancer cells in a time- and concentration-dependent manner. These results showed that combined therapy with low concentrations were detected to be more effective than that with high-concentration alone drug treatment. 

Keywords

References

1. Abramson VG, Lehmann BD, Ballinger TJ, Pietenpol JA. Subtyping of triple-negative breast cancer: implications for therapy. Cancer 2015; 121: 8-16.
2. Lyons TG. Targeted therapies for triple-negative breast cancer. Curr Treat Options Oncol 2019; 20: 82.
3. Canfield K, Feng W, Kurokawa M. Metabolic regulation of apoptosis in cancer. Int Rev Cell Mol Biol 2016; 327: 43-87. 
4. Parton M, Dowsett M, Smith, I. Studies of apoptosis in breast cancer. BMJ 2001; 322: 1528–1532
5. Harbeck N, Gnant M. Breast cancer. Lancet 2017; 18: 1134-1150. 
6. Bayat Mokhtari R, Homayouni TS, Baluch N, Morgatskaya E, Kumar S, Das B et al. Combination therapy in combating cancer.  Oncotarget 2017; 8: 38022-38043.
7. Eroglu O, GÇ Esin, Kaya H, Celen M, Karabicici M, Karacoban E. Investigation of methylation profiles of TP53, Caspase 9, Caspase 8, Caspase 3 genes treated with DNA methyl transferase ınhibitor (DNMTi) zebularine (ZEB) and caffeic acid phenethyl ester (CAPE) on MCF-7 and MDA-MB-231 breast cancer cell line. J Cancer Ther 2019; 10: 69-85. 
8. Frangione ML, Lockhart JH, Morton DT, Pava LM, Blanck G. Anticipating designer drug-resistant cancer cells. Drug Discov Today 2015; 20:790-793.
9. Chiappinelli KB, Zahnow CA, Ahuja N, Baylin SB. Combining epigenetic and ımmunotherapy to combat cancer. Cancer Res 2016; 76 :1683-1689. 
10. Jerusalem G, Collignon J, Schroeder H, Lousberg L. Triple-negative breast cancer: treatment challenges and solutions. Breast Cancer (Dove Med Press) 2016;8: 93-107.
11. New M, Olzscha H, La Thangue NB. HDAC inhibitor-based therapies: can we interpret the code? Mol Oncol 2012; 6: 637-656.
12. Fournel M, Bonfils C, Hou Y, Yan PT, Trachy-Bourget MC, Kalita A, et al. MGCD0103, a novel isotype-selective histone deacetylase inhibitor, has broad spectrum antitumor activity in vitro and in vivo. Mol Cancer Ther 2008; 7: 759–768. 
13. Boumber Y, Younes A, Garcia-Manero G. Mocetinostat (MGCD0103): A review of an isotype-specific histone deacetylase inhibitor. Expert Opin Investig Drugs 2011; 20: 823–829. 
14. Siu LL, Pili R, Duran I, Messersmith WA, Chen EX, Sullivan R, et al. Phase I study of MGCD0103 given as a three-times-per-week oral concentration in patients with advanced solid tumors. J Clin Oncol 2008;26: 1940–1947. 
15. Park DJ, Stoehlmacher J, Zhang W, Tsao-Wei D, Groshen S, Lenz HJ. Thymidylate synthase gene polymorphism predicts response to capecitabine in advanced colorectal cancer. Int J Colorectal Dis 2002;17: 46-49. 
16. Eroğlu O, Kaya H, Çelik E, Korkut E. Nizam N. Triple effect of doxorubicin, 5-fluorouracil, propranolol on cell survival on MCF-7 breast cancer cell line. J Biosci Med (Irvine) 2019; 7: 74-85.
17. Mendis AS, Thabrew I, Samarakoon SR, Tennekoon KH. Modulation of expression of heat shock proteins and apoptosis by Flueggea leucopyrus (Willd) decoction in three breast cancer phenotypes. BMC Complement Altern Med 2015; 9; 404. 
18. Li J, Guo Y, Duan L, Hu X, Zhang X, Hu J, et al. AKR1B10 promotes breast cancer cell migration and invasion via activation of ERK signaling. Oncotarget 2017; 16; 33694-33703.
19. Gottesman MM, Fojo T, Bates SE. Multidrug resistance in cancer: Role of ATP-dependent transporters. Nat Rev Cancer 2002;2: 48-58. 
20. Daniele G, Gallo M, Piccirillo MC, Giordano P, D’Alessio A, Del Giudice A, et al. Pharmacokinetic evaluation of capecitabine in breast cancer. Expert Opin Drug Metab Toxicol 2013; 9: 225–235. 
21. Gligorov J, Doval D, Bines J, Alba E, Cortes P, Pierga JY et al. Maintenance capecitabine and bevacizumab versus bevacizumab alone after initial first-line bevacizumab and docetaxel for patients with HER2-negative metastatic breast cancer (IMELDA): A randomised, open-label, phase 3 trial. Lancet Oncol. 2014; 15: 1351–1360. 
22. Budillon A, Di Gennaro E, Bruzzese F, Rocco M, Manzo G and Caraglia M. Histone deacetylase inhibitors: a new wave of molecular targeted anticancer agents. Recent Pat Anticancer Drug Discov 2007; 2:119–134. 
23. Lee HZ, Kwitkowski VE, Del Valle PL, Ricci MS, Saber H, Habtemariam BA, et al. FDA approval: Belinostat for the treatment of patients with relapsed or refractory peripheral T-cell lymphoma. Clin Cancer Res 2015; 15: 2666-2670. 
24. Raha P, Thomas S, Munster PN. Epigenetic modulation: a novel therapeutic target for overcoming hormonal therapy resistance. Epigenomics 2011; 3: 451–470. 
25. Candelaria M, Cruz-Hernandez E, Taja-Chayeb L, Perez-Cardenas E, Trejo-Becerril C, Gonzalez-Fierro A et al. DNA methylation-independent reversion of gemcitabine resistance by hydralazine in cervical cancer cells. PLoS One 2012; 7: 29181. 
26. Baradaran PC, Mohammadi A, Mansoori B, Baradaran SC, Baradaran B. Growth inhibitory effect of Scrophularia oxysepala extract on mouse mammary carcinoma 4T1 cells in vitro and in vivo systems. Biomed Pharmacother 2017; 85: 718-724. 
27. Zhang SH, Zhang H, He HW, Li L, Li XQ, Zhang YP, et al. Lidamycin up-regulates the expression of thymidine phosphorylase and enhances the effects of capecitabine on the growth and pulmonary metastases of murine breast carcinoma. Cancer Chemother Pharmacol. 2013; 72: 777-788. 
28. Guichard SM, Macpherson JS, Mayer I, Reid E, Muir M, Dodds M, et al. Gene expression predicts differential capecitabine metabolism, impacting on both pharmacokinetics and antitumour activity. Eur J Cancer 2008; 44: 310-317. 
29. Kell J. Drug evaluation: MGCD-0103, a histone deacetylase inhibitor for the treatment of cancer. Curr Opin Investig Drugs 2007; 8: 485-492. 
30. Lopez G, Braggio D, Zewdu A, Casadei L, Batte K, Bid HK, et al. Mocetinostat combined with gemcitabine for the treatment of leiomyosarcoma: Preclinical correlates. PLoS One 2017 29; 12: 0188859. 
31. Hontecillas-Prieto L, Flores-Campos R, Silver A, Álava E, Hajji N, García-Domínguez DJ. Synergistic enhancement of cancer therapy using HDAC ınhibitors: Opportunity for clinical trials. Front Genet 2020; 11: 578011.
32. Li J, Yu K, Pang D, Wang C, Jiang J, Yang S, et al. Adjuvant capecitabine with docetaxel and cyclophosphamide plus epirubicin for triple-negative breast cancer (CBCSG010): An open-label, randomized, multicenter, phase III trial. J Clin Oncol 2020; 38: 1774–1784O’Shaughnessy J. Capecitabine and docetaxel in advanced breast cancer: analyses of a phase III comparative trial. Oncology 2002; 16: 17–22.
33. O’Shaughnessy J. Capecitabine and docetaxel in advanced breast cancer: analyses of a phase III comparative trial. Oncology (Williston Park) 2002; 16: 17–22. 
34. Zhang Q, Sun M, Zhou S, Guo B. Class I HDAC inhibitor mocetinostat induces apoptosis by activation of miR-31 expression and suppression of E2F6. Cell Death Discov 2016; 2:16036.
35. Madden R, Kosari S, Peterson GM, Bagheri N, Thomas J. Lapatinib plus capecitabine in patients with HER2-positive metastatic breast cancer: A systematic review. Int J Clin Pharmacol Ther 2018; 56: 72-80. 
36. Leone A, Roca MS, Ciardiello C, Terranova-Barberio M, Vitagliano C, Ciliberto G, et al. Vorinostat synergizes with EGFR inhibitors in NSCLC cells by increasing ROS via up-regulation of the major mitochondrial porin VDAC1 and modulation of the c-Myc-NRF2-KEAP1 pathway. Free Radic Biol Med 2015; 89: 287–299
37. Seo J, Min SK, Park HR, Kim DH, Kwon MJ, Kim LS et al. Expression of histone deacetylases HDAC1, HDAC2, HDAC3, and HDAC6 in ınvasive ductal carcinomas of the breast. J Breast Cancer 2014; 17: 323-331. 
38. Wisniewska-Jarosinska M, Sliwinski T, Kasznicki J, Kaczmarczyk D, Krupa R,  Bloch K, et al. Cytotoxicity and genotoxicity of capecitabine in head and neck cancer and normal cells. Mol Biol Rep 2011; 38: 3679-3688. 
39. Fulda S. Modulation of TRAIL-induced apoptosis by HDAC inhibitors. Curr Cancer Drug Targets 2008; 8: 132-140. 
40. Chen M, Chen J, Peng X, Xu Z, Shao J, Zhu Y, et al. The contribution of keratinocytes in capecitabine-stimulated hand-foot-syndrome. Environ Toxicol Pharmacol 2017; 49: 81-88. 
41. Li M, Zhang N, Li M, Capecitabine treatment of HCT-15 colon cancer cells induces apoptosis via mitochondrial pathway. Trop J Pharm Res 2017; 16: 1529-1536.
42. Liao B, Sun Q, Yuan Y, Yin Y, Qiao J, Jiang P. Histone deacetylase inhibitor MGCD0103 causes cell cycle arrest, apoptosis, and autophagy in liver cancer cells. J Cancer 2020; 29: 1915-1926.
Iranian Journal of Basic Medical Sciences
Volume 24, Issue 11
November 2021
Pages 1515-1522

Files

Share

How to cite

Statistics