Background. Curcumin (CUR), a natural phenolic compound, has been recently reported to exert antitumor actions in variety of cancers; however, the exact mechanism(s) is not clear. In this study we investigated whether CUR could inhibit Ras/MAPK pathway and enhance mitomycin C (MMC) cytotoxicity in T24 bladder cancer cells. Methods. T24 cells were cultured with different concentrations of CUR (5, 10, 20 μM) alone or combined with 10 μg/ml MMC. At the end of 72 h culture, cell viability was assessed by MTT assay; apoptosis by flow cytometry; total Ras and ERK1/2 by immunohistochemistry and western blotting. Results. In comparison to cells exposed to MMC alone, cells treated with combined MMC and either 10 or 20 μM CUR showed reduced cell proliferation, disrupted morphological appearance, and increased subG0/G1 apoptotic events. This inhibition was associated with marked reduction of Ras and ERK1/2 expression. Likewise, cells treated with 10 or 20 μM CUR alone showed significant inhibition, while the effect of 5 μM was less obvious. Conclusion. Resistance of T24 cells to cytotoxic effect of MMC is dependent, at least partially, on Ras/ERK activation. CUR at concentrations of 10 and 20 μM in combination with low dose MMC induced toxic synergism in T24 cells. Clinical translation of this experimental study may be reasonable in light of wide safety margin and availability of CUR.
curcumin, mitomycin C, Ras siRNA, ERK, cancer bladder
Hurst CD, Alder O, Platt FM, Droop AD, Stead LF, Burns JE, Burghel GJ, Jain S, Klimczak LJ, Lindsay H, Roulson J, Taylor CF, Thygesen H, Cameron AJ, Ridley AJ, Mott HR, Gordenin DA, Knowles MA. The genomic landscape of non-muscle-invasive bladder cancer: implications for molecular
classification and treatment. Cancer Res, 2016; 76: LB–323.
Birare N, Lwaleed BA, Cooper AJ: Multidrug resistance in a urothelial cancer cell line after 1-hour mitomycin C exposure. J Urol. 2009; 182: 2472–76.
Xu B, Sun Y, and Singh SV: Mechanism of resistance to mitomycin C in a human bladder cancer cell line. Zhonghua Zhong Liu Za Zhi. 1995; 17: 343–6.Youssef et al. 2018 Curcumin enhances cytotoxicity of mitomycin C
Au JL, Badalament RA, Wientjes MG, Young DC, Warner JA, Venema PL, Pollifrone DL, Harbrecht JD, Chin JL, Lerner SP, Miles BJ. Methods to improve efficacy of intravesical mitomycin C. results of a randomized phase III trial. J Natl Cancer Inst. 2001; 93:597.
Shinohara N, Koyanagi T. Ras signal transduction in carcinogenesis and progression of bladder cancer: molecular target for treatment? Urol Res. 2002; 30: 273–81.
Parada LF, Tabin CJ, Shih C, Weinberg RA. Human EJ bladder carcinoma oncogene is homologue of Harvey sarcoma virus ras gene. Nature. 1982; 297: 474–8
Johnson GL, Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science, 2002; 298:1911–12
Maertens O, Cichowski K. An expanding role for RAS GTPase activating proteins (RAS GAPs) in cancer. Adv Biol Reg. 2014; 55:1–14.
Boulalas I, Zaravinos A, Karyotis I, Delakas D, Spandidos DA. Activation of RAS family genes in urothelial carcinoma. J Urol. 2009; 181: 2312–19.
Johne A, Roots I, and Brockmöller J. A single nucleotide polymorphism in the human H-ras proto-oncogene determines the risk of urinary bladder cancer. Cancer Epidemiol Biomarkers Prev. 2003; 12: 68–70.
Chang F, Steelman LS, Lee JT, Shelton JG, Navolanic PM, Blalock WL, Franklin RA, and McCubrey JA. Signal transduction mediated by the Ras/Raf/MEK/ERK pathway from cytokine receptors to transcription factors: potential targeting for therapeutic intervention. Leukemia. 2003; 17:
Samatar AA, and Poulikakos PI: Targeting RAS–ERK signalling in cancer: promises and challenges. Nature Rev Drug Dis, 2014; 13: 928–42.
Nelson KM, Dahlin JL, Bisson J, Graham J, Pauli GF, Walters MA. The essential medicinal chemistry of curcumin: miniperspective. J Med Chem. 2017; 60: 1620–1637.
Kasi PD, Tamilselvam R, Skalicka-Woźniak K, Nabavi SF, Daglia M, Bishayee A, Pazoki-Toroudi H, Nabavi SM. Molecular targets of curcumin for cancer therapy: an updated review. Tumour Biol. 2016;37:13017-13028.
Shehzad A, Lee YS. Molecular mechanisms of curcumin action: signal transduction. Biofactors. 2013; 39:27–36.
Ono M, Higuchi T, Takeshima M, Chen C, Nakano S. Differential anti-tumor activities of curcumin against Ras- and Src-activated human adenocarcinoma cells. Biochem Biophys Res Commun. 2013; 436:186-91.
Sharaf Eldin O, Fouda AM, Youssef AR, Hamilton P, Maxwell P, and Williamson KE. Reduction of mitomycin C resistance in human bladder cancer T24 cells by knocking-down Ras oncogene. Cancer Drug Resist. 2018; 1:59-71.
Vasquez JL, Gehl J, and Hermann GG: Electroporation enhances mitomycin C cytotoxicity on T24 bladder cancer cell line: a potential improvement of intravesical chemotherapy in bladder cancer. Bioelectrochemistry. 2012; 88: 127–33.
Florento L, Matias R, Tuaño E, Santiago K, dela Cruz F, Tuazon A. Comparison of Cytotoxic Activity of Anticancer Drugs against Various Human Tumor Cell Lines Using In Vitro Cell-Based Approach. Int J Biomed Sci. 2012;8(1):76-80.
Dahab GM, Kheriza M, El-Beltagi HM, Fouda AM, and Sharaf El-Din O: Digital quantification of fibrosis in liver biopsy sections: description of a new method by Photoshop software. J Gastroenterol Hepatol. 2004; 19: 78–85.
Rizzardi AE, Johnson AT, Vogel RI, Pambuccian SE, Henriksen J, Skubitz AP, Metzger GJ, and Schmechel SC: Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring. Diagn Pathol. 2012; 7: 42.
Oxford G and Theodorescu D: The role of Ras superfamily proteins in bladder cancer progression. J Urol. 2003; 170: 1987–93.
Maertens O, Cichowski K. An expanding role for RAS GTPase activating proteins (RAS GAPs) in cancer. Advances in Biological Regulation. 2014;55:1–14.
Das T, Sa G, Saha B, Das K. Multifocal signal modulation therapy of cancer: Ancient weapon, modern targets. Mol Cell Biochem. 2010; 336: 85-95.
Hancock JF. RAS protein: Different signals from different locations. Nat Rev Mol Cell Biol. 2003; 4: 373-384.
Manuel M. Paz, Xu Zhang, Jun Lu, Holmgren A. A new mechanism of action for the anticancer drug mitomycin C: mechanism-based inhibition of thioredoxin reductase. Chemical Res Toxicol. 2012; 25: 1502-1511.
Stanslas J, Wong CC, Sagineedu SR, Sidik S, Sumon SH, Phillips R, Lajis NH. Mechanism of resistance to a semisynthetic anticancer andrographolide derivative: Possible involvement of Ras-MAPK signaling pathway. Mol Cancer Ther. 2013; 12: C147.
Revalde JL, Li Y, Hawkins BC, Rosengren RJ, Paxton JW. Heterocyclic cyclohexanone monocarbonyl analogs of curcumin can inhibit the activity of ATP-binding cassette transporters in cancer multidrug resistance. Biochem Pharmacol. 2015; 93:305–17.
Zhou QM, Wang XF, Liu XJ, Zhang H, Lu YY, Huang S. Curcumin improves MMC-based chemotherapy by simultaneously sensitising cancer cells to MMC and reducing MMC-associated side-effects. Eur J Cancer. 2011; 47: 2240–2247.
Rashmi R, Kumar S, Karunagaran D. Ectopic expression of Hsp70 confers resistance and silencing its expression sensitizes human colon cancer cells to curcumin-induced apoptosis. Carcinogenesis. 2004;25:179–87.
Johnson GL and Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science. 2002; 298:1911–12
Chang F, Steelman LS, Lee JT, Shelton JG, Navolanic Youssef et al. 2018
Curcumin enhances cytotoxicity of mitomycin C
PM, Blalock WL, Franklin RA, and McCubrey JA. Signal transduction mediated by the Ras/Raf/MEK/ERK pathway from cytokine receptors to transcription factors: potential targeting for therapeutic intervention. Leukemia. 2003; 17: 1263–93