Document Type : Original Article
Authors
Department of Biology, Faculty of Sciences, Urmia University, Urmia, Iran
Abstract
Inflammation is an initial response of the body to infection and relationship between inflammation and cancer has been established. Nuclear factor kappa B (NF-κB) is a central factor in inflammation and its activity contributes to tumor progression and apoptosis prevention consequently leading to cancer promotion. As a result, NF-κB inhibitors can cause apoptosis. In this study, the effect of mesalazine as a NF-κB inhibitor on growth and apoptosis of K562 cells has been investigated. The K562 cells were first cultured in RPMI-1640 medium containing 10.00% fetal bovine serum. After that, they were treated for 72 hr with different concentrations of mesalazine (20.00, 40.00, 60.00 and 80.00 μM mL-1). The MTT assay was used to evaluate cell viability. Hoechst staining and RT-PCR of apoptosis related genes (Bcl-2 and Bax) were carried out to illustrate apoptosis induction and immunocytochemistry was performed to investigate changes in c-Myc protein level. According to the results of MTT assay, all of applied mesalazine concentrations decreased K562 cells viability. Hoechst staining showed that the fragmented nuclei increased indicating apoptosis induction. Immuno-cytochemical results showed that mesalazine decreased c-Myc in treated cells. The RT-PCR results also showed an increase in Bax and a decrease in Bcl-2 expressions in mesalazine-treated cells. As the results suggest, mesalazine reduces cell viability by inducing apoptosis in K562 cell line; therefore, it can be used as a candidate for the leukemia treatment.
Keywords
- Gursoy O, Kinik O. Probiotics: a new popular option for cancer inhibition. Int J Dairy Sci 2006; 1(1): 100-103.
- Quintás-Cardama A, Cortes JE. Chronic myeloid leukemia: diagnosis and treatment. Mayo Clin Proc 2006; 81(7): 973-988.
- Han SI, Kim YS, Kim TH. Role of apoptotic and necrotic cell death under physiologic conditions. BMB Rep 2008; 41(1): 1-10.
- Letai A. Apoptosis and cancer. Annu Rev Cancer Biol 2017; 1: 275-294
- Chen L, Deng H, Cui H, et al. Inflammatory responses and inflammation-associated diseases in organs. Oncotarget 2017; 9(6): 7204-7218.
- TakPP, FiresteinGS. NF-kappaB: a key role in inflammatory diseases. J Clin Invest. 2001; 107(1):
7-11. - Karagozian R, Burakoff R. The role of mesalamine in the treatment of ulcerative colitis. Ther Clin Risk Manag 2007; 3(5): 893-903.
- Bos CL, Diks SH, Hardwick JCH, et al. Protein phosphatase 2A is required for mesalazine-dependent inhibition of Wnt/beta-catenin pathway activity. Carcinogenesis 2006; 27(12): 2371-2382.
- van Meerloo J, Kaspers GJL, Cloos J. Cell sensitivity assays: the MTT assay. Methods Mol Biol 2011; 731: 237-245.
- Bijnsdorp IV, Giovannetti E, Peters GJ. Analysis of drug interactions. Methods Mol Biol 2011; 731: 421-434.
- Allen S, Sotos J, Sylte MJ, et al. Use of Hoechst 33342 staining to detect apoptotic changes in bovine mononuclear phagocytes infected with Mycobacterium avium paratuberculosis. Clin Diagn Lab Immunol 2001; 8(2): 460-464.
- Johansson BG. Agarose gel electrophoresis. Scand J Clin Lab Invest 1972; 124: 7-19.
- Pazhang Y, Ahmadian S, Javadifar N, et al. COX-2 and survivin reduction may play a role in berberine-induced apoptosis in human ductal breast epithelial tumor cell line. Tumour Biol 2012; 33(1): 207-214.
- Liu T, Zhang L, Joo D, et al. NF-κB signaling in inflammation. Signal Transduct Target Ther 2017, 2: 17023. doi: 10.1038/sigtrans.2017.23.
- Xia L, Tan S, Zhou Y, et al. Role of the NFκB-signaling pathway in cancer. Onco Targets Ther 2018, 11:
2063-2073. - Hoesel B, Schmid JA. The complexity of NF-κB signaling in inflammation and cancer. Mol Cancer 2013; 12: 86. doi:10.1186/1476-4598-12-86
- Miller SC, Huang R, Sakamuru S, et al. Identification of known drugs that act as inhibitors of NF-kappa B signaling and their mechanism of action. Biochem Pharmacol 2010; 79(9): 1272-1280.
- Sekino N, Kano M, Matsumoto Y, et al. Antitumor effects of metformin are a result of inhibiting nuclear factor kappa B nuclear translocation in esophageal squamous cell carcinoma. Cancer Sci 2018; 109 (4): 1066-1074.
- Sena P, Mancini S, Benincasa M, et al. Metformin induces apoptosis and alters cellular responses to oxidative stress in Ht29 colon cancer cells: preliminary findings. Int J Mol Sci 2018; 19(5): 1478. doi: 10.3390/ ijms19051478.
- Ni H, Zhao W, Kong X, et al. NF-kappa B modulation is involved in celastrol induced human multiple myeloma cell apoptosis. PLoS One 2014; 9(4): e95846. doi: 10.1371/journal.pone.0095846
- Wang Z, Zhai Z, Du, X. Celastrol inhibits migration and invasion through blocking the NF-κB pathway in ovarian cancer cells. Exp Ther Med 2017; 14(1): 819-824.
- Yamamoto Y, Yin MJ, Lin KM, et al. Sulindac inhibits activation of the NF-κB pathway. J Biol Chem 1999; 274(38): 27307-27314.
- Reinacher-Schick A, Schoeneck A, Graeven U, et al. Mesalazine causes a mitotic arrest and induces caspase-dependent apoptosis in colon carcinoma cells. Carcinogenesis 2003; 24(3): 443-451.
- Wang LW, Li ZS, Zou DW, et al. Metformin induces apoptosis of pancreatic cancer cells. World J Gastroenterol 2008; 14(47): 7192-7198.
- Elbadawy M, Usui T, Yamawaki H, et al. Emerging roles of c-Myc in cancer stem cell-related signaling and resistance to cancer chemotherapy: a potential therapeutic target against colorectal cancer. Int J Mol Sci 2019; 20(9): 2340. doi: 10.3390/ijms20092340.
- Stolfi C, Pellegrini R, Franze E, et al. Molecular basis of the potential of mesalazine to prevent colorectal cancer. World J Gastroenterol 2008; 14(28): 4434-4439.
- Klapproth K, Sander S, Marinkovic D, et al. The IKK2/NF-{kappa} B pathway suppresses MYC-induced lymphomagenesis. Blood 2009; 114(12): 2448-2458.
)