Document Type : Original Article

Authors

1 Assistant Professor Department of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

2 Department of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.

3 Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Abstract

Introduction and Aim: Colorectal cancer is one of the most common cancers. Epigenetic change has been considered by many scientists as a therapeutic target. Hyper acetylation of chromatin components by sodium butyrate can alter gene regulation. This study aims to investigate the effects of sodium butyrate on Bax and Bcl-2 gene expression.
Methods: Caco-2 cell line was treated with different concentrations of sodium butyrate (25 mM to 150 mM) based on IC50 concentration in two time periods of 24 hours and 48 hours. Bax and Bcl-2 gene expression were measured by qReal-Time PCR technique and Bcl2/Bax ratio was evaluated.
Results: The results showed that sodium butyrate increased the expression of Bax gene and decreased the expression of Bcl-2 gene in treated cells compared to the control group, which was statistically significant (p < /em> <0.05), and 25 mM was selected as the most effective dose after 48 hours of treatment. Also, the Bcl-2/Bax ratio at the same concentration showed a significant decrease
Conclusion: Sodium butyrate induces apoptosis in cancer cells by reducing the expression ratio of Bcl-2/Bax. It can be used as a therapeutic target but needs further investigation.

Keywords

Main Subjects

Amiri, N., Forouzesh, F., Nazemalhosseini- Mojarad, E., Shabani, M. 2020. The effect of sodium butyrate as a histone deacetylase inhibitor on the gene expression of Bid in HT-29 human colorectal cancer cell line. Medical Science Journal of Islamic Azad University, Tehran Medical Branch, 30(1): 40-50.
Arvelo, F., Sojo, F. & Cotte, C. 2015. Biology of colorectal cancer. Ecancermedicalscience: 9.
Chirakkal, H., Leech, S., Brooks, K., Prais, A., Waby, J. & Corfe, B. 2006. Upregulation of BAK by butyrate in the colon is associated with increased Sp3 binding. Oncogene, 25: 7192-7200.
Chopin, V., Toillon, R. A., Jouy, N. & Bourhis, X. L. 2002. Sodium butyrate induces P53‐independent, Fas‐mediated apoptosis in MCF‐7 human breast cancer cells. British journal of pharmacology, 135: 79-86.
Chueh, A. C., Tse, J. W., Togel, L. & Mariadason, J. M. 2015. Mechanisms of histone deacetylase inhibitor-regulated gene expression in cancer cells. Antioxidants & redox signaling, 23: 66-84.
Davie, J. R. 2003. Inhibition of histone deacetylase activity by butyrate. The Journal of nutrition, 133: 2485S-2493S.
Dejakam, A., Dejakam, A. and Hekmat, A. 2020. Evaluating of bdnf expression in blood cells of opium recovering patients with a new treatment method: A molecular marker. Research in Karyotic Cell &Tissue, 1(1): 16-25.
Del Poeta, G. Venditti, A. Del Principe, M.I., et al. 2003. Amount of spontaneous apoptosis detected by Bax/Bcl-2 ratio predicts outcome in acute myeloid leukemia (AML). Blood, 101:2125–2131.
Esteller, M. 2008. Epigenetics in cancer. New England Journal of Medicine, 358: 1148-1159.
Farkas, R. Pozsgai, E., Bellyei, S.Z., Cseke, L., Szigeti, A., Vereczkei, A., Marton, S., Mange, L., Horvath, O.P., Papp, A. 2011. Correlation between tumorassociated proteins and response to neoadjuvant treatment in patients with advanced squamous-cell esophageal cancer. Anticancer research. 31(5):1769-75.
Forouzesh,  F., Talebi , S., Shahbazi, N. 2018. Anticancer effects of sodium butyrate on human colon cancer Caco2 cell line. Journal of Fasa University of Medical Sciences, 8: 735-745.
Forouzesh, F., Ghiaghi, M., & Rahimi, H. 2020. Effect of sodium butyrate on HDAC8 mRNA expression in colorectal cancer cell lines and molecular docking study of LHX1 - sodium butyrate interaction. EXCLI Journal, 19: 1038-1051.
Fraga,M. F., Ballestar, E., Villar-Garea,  A., Boix-Chornet, M., Espada, J., Schotta,  G., Bonaldi, T., Onaldi, T., Haydon,  C., Ropero, S. & Petrie, K. 2005. Loss of acetylation at Lys16 and trimethylation at Lys20 of histone H4 is a common hallmark of human cancer. Nature genetics, 37: 391-400.
Gum, J., Kam W., Byrd, J., Hicks, J., Sleisenger, M. & Kim, Y. 1987. Effects of sodium butyrate on human colonic adenocarcinoma cells. Induction of placental-like alkaline phosphatase. Journal of Biological Chemistry, 262: 1092-1097.
Hekmat, A. and Saboury, A.A. 2019. Structural effects of the syntheticcobalt–manganese-zinc ferrite nanoparticles (Co0.3Mn0.2Zn0.5 Fe2O4NPs) on DNA and its antiproliferative effect on t47dcells. BioNanoScience, 9(4): 821-832.
Huerta, S., Goulet, E. J. & Livingston, E. H. 2006. Colon cancer and apoptosis. The American Journal of Surgery, 191: 517-526.
Jia, Y. & Guo, M. 2013. Epigenetic changes in colorectal cancer. Chinese journal of cancer, 32: 21.
Khan, S. & Jena, G. 2014. Protective role of sodium butyrate, a HDAC inhibitor on beta-cell proliferation, function and glucose homeostasis through modulation of p38/ERK MAPK and apoptotic pathways: study in juvenile diabetic rat. Chemico-biological interactions, 213: 1-12.
Khaw, S.L., Mérino, D., Anderson, M.A., Glaser, S.P., Bouillet, P., Roberts, A.W. & Huang, D.C.S.
2014. Both leukaemic and normal peripheral B lymphoid cells are highly sensitive to the selective Pharmacological inhibition of prosurvival Bcl-2 with ABT-199. Leukemia,28:1207–1215.
Kilm, Y. S., Tsao, D., Siddiqui , B., Whitehead, J. S., Arnstein , P., Bennett , J. & Hicks, J. 1980. Effects of sodium butyrate and dimethylsulfoxide on biochemical properties of human colon cancer cells. Cancer, 45: 1185-1192.
Koff, J. L., Ramachandiran, S. & Bernal-Mizrachi, L. 2015. A time to kill: targeting apoptosis in cancer. International journal of molecular sciences, 16: 2942-2955.
Kulsoom, B., Shamsi, T.S., Afsar, N.A., Memon, Z., Ahmed, N., Hasnain, S.N. 2018. Bax, Bcl-2, and Bax/Bcl-2 as prognostic markers in acute myeloid leukemia: are we ready for Bcl-2-directed therapy? Cancer Management and Research,10: 403-416.
Mariadason, J. M. 2008. HDACs and HDAC inhibitors in colon cancer. Epigenetics, 3: 28-37.
Qian, H.-R., Shi, Z.Q., Zhu, H.P., Gu, L.H., Wang, X.F. & Yang, Y. 2017. Interplay between apoptosis and autophagy in colorectal cancer. Oncotarget, 8: 62759.
Mashayekhi,  A., Forouzesh, F.,  and  Mashayekhi, M. 2021. Promotion of Extrinsic Apoptosis Pathway in HCT‐116 Human Colorectal Cancer Cell Line by Sodium Butyrate as Histone Deacetylase Inhibitor. Iran Red Crescent Medical Journal. 23(1):e190.
Ruemmele, F., Schwartz, S., Seidman, E., Dionne, S., Levy, E. & Lentze, M. 2003. Butyrate induced Caco-2 cell apoptosis is mediated via the mitochondrial pathway. Gut, 52: 94-100.
Salimi, V., Shahsavari, Z., Safizadeh, B., Hosseini, A., Khademian, N., and Tavakoli-Yaraki, M. 2017. Sodium butyrate promotes apoptosis in breast cancer cells through reactive oxygen species (ROS) formation and mitochondrial impairment. Lipids Health Dis, 16: 208.
Shadmani, F. K., Ayubi, E., Khazaei, S., Sani, M., Hanis, S. M., Khazaei, S., Soheylizad, M., & Mansori, K. 2017. Geographic distribution of the incidence of colorectal cancer in Iran: a population-based study. Epidemiology and health, 39: e2017020.
Sharma, S., Kelly, T. K., & Jones, P. A. 2010. Epigenetics in cancer. Carcinogenesis, 31: 27-36.
Spurling, C. C., Godman, C. A., Noonan, E. J., Rasmussen, T. P., Rosenberg, D. W.,. & Giardina, C. 2008. HDAC3 overexpression and colon cancer cell proliferation and differentiation. Molecular Carcinogenesis: Published in cooperation with the University of Texas MD Anderson Cancer Center, 47: 137-147.
Stunkel, W., Peh, B. K., Tan, Y. C., Nayagam, V. M., Wang, X., Salto-Tellez, M., Ni, B., Entzeroth, M., & Wood, J. 2007. Function of the SIRT1 protein deacetylase in cancer. Biotechnology Journal: Healthcare Nutrition Technology, 2: 1360-1368.
Wang, W., Fang, D., Zhang, H., Xue, J., Wangchuk, D., Du, J., Jiang, L. 2020. Sodium Butyrate Selectively Kills Cancer Cells and Inhibits Migration in Colorectal Cancer by Targeting Thioredoxin-1. OncoTargets and Therapy 13: 4691-4704.
 Watson, A. 2004. Apoptosis and colorectal cancer. Gut, 53: 1701-1709.
West, A. C. & Johnstone, R. W. 2014. New and emerging HDAC inhibitors for cancer treatment. The Journal of clinical investigation, 124:30-39.
Wilson, A. J., Byun, D.S., Popova, N., Murray, L. B., Litalien, K., Sowa, Y., Aramgo, D., Velcich, A., Augenlicht, L. H., & Mariadason, J. M. 2006. Histone deacetylase 3 (HDAC3) and other class I HDACs regulate colon cell maturation and p21 expression and are deregulated in human colon cancer. Journal of Biological Chemistry, 281: 13548-13558.
Xu, F., Zhang, K., & Grunstein, M. 2005. Acetylation in histone H3 globular domain regulates gene expression in yeast. Cell, 121: 375-385.
Yan, W., & Guo, M. 2015. Epigenetics of colorectal cancer. Methods in Molecular Biology, 1238:405-24
Zhu, P., Martin, E., Mengwasser, J., Schlag, P., Janssen, K.P. & Gottlicher, M. 2004. Induction of HDAC2 expression upon loss of APC in colorectal tumorigenesis. Cancer cell, 5: 455-463.