Glioblastoma is the most aggressive and fatal form of brain cancer. Despite new advancements in treatment, the desired outcomes have not been achieved. Temozolomide (TMZ) is the first-choice treatment for the last two decades and has improved survival rates. Emerging studies have shown that targeting epigenetics in glioblastoma can be beneficial when combined with clinically used treatments. Trichostatin A (TSA), a histone deacetylase inhibitor, has anti-cancer properties in various cancers. No data concerning the TMZ and TSA relationship was shown previously in glioblastoma therefore, we aimed to determine the likely therapeutic effect of the TMZ and TSA combination in glioblastoma. The T98G and U-373 MG, glioblastoma cell lines, were used in this study. TMZ and TSA cytotoxicity and combination index were performed by MTT assay. The expression of DNA repair genes (MGMT, MLH-1, PMS2, MSH2 and MSH6) was detected using RT-PCR. One-way analysis of variance (ANOVA) was used for statistical analysis. Combination index calculations revealed antagonistic effects of TMZ and TSA in terms of cytotoxicity. Antagonistic effects were more apparent in the T98G cell line, which is expressing MGMT relatively higher. MGMT and DNA Mismatch Repair (MMR) genes were upregulated in the T98G cell line, whereas downregulated in the U373-MG cell lines under TMZ and TSA combination treatment. It is concluded that MGMT might be playing a more active part than MMR genes in TMZ resistance to TMZ and TSA antagonism. This is the first study elucidating the TMZ and TSA relationship in cancer cell lines.
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Ostrom QT, et al. CBTRUS Statistical Report: primary brain and other Central Nervous System Tumors diagnosed in the United States in 2015–2019. Neuro Oncol. 2022;24(5):v1–v. https://doi.org/10.1093/neuonc/noac202.
Tykocki T, Eltayeb M. Ten-year survival in glioblastoma. A systematic review. J Clin Neurosci. 2018;54:7–13. https://doi.org/10.1016/j.jocn.2018.05.002.
Louis DN, et al. The 2021 WHO classification of tumors of the central nervous system: a summary. Neuro Oncol. 2021;23(8):1231–51. https://doi.org/10.1093/neuonc/noab106.
Silvani A. New perspectives: glioma in adult patients. Tumori J. p. Mar. 2023;030089162311597. https://doi.org/10.1177/03008916231159716.
Ståhl P, Henoch I, Smits A, Rydenhag B, Ozanne A. Quality of life in patients with glioblastoma and their relatives. Acta Neurol Scand. 2022;146(1):82–91. https://doi.org/10.1111/ane.13625.
Shi J, Dong B, Zhou P, Guan W, Peng Y. Functional network analysis of gene-phenotype connectivity associated with temozolomide. Oncotarget. 2017;8(50):87554–67. https://doi.org/10.18632/oncotarget.20848.
Liu EK, Sulman EP, Wen PY, Kurz SC. Novel therapies for Glioblastoma. Curr Neurol Neurosci Rep. 2020;20(7). https://doi.org/10.1007/s11910-020-01042-6.
Lee P, et al. Mechanisms and clinical significance of histone deacetylase inhibitors: epigenetic glioblastoma therapy. Anticancer Res. 2015;35(2):615–25.
Kunadis E, Lakiotaki E, Korkolopoulou P, Piperi C. Targeting post-translational histone modifying enzymes in glioblastoma. Pharmacol Ther. 2021;220:107721. https://doi.org/10.1016/j.pharmthera.2020.107721.
Zhang Y, Carr T, Dimtchev A, Zaer N, Dritschilo A, Jung M. Attenuated DNA damage repair by trichostatin a through BRCA1 suppression. Radiat Res. 2007;168(1):115–24. https://doi.org/10.1667/RR0811.1.
Perla A, et al. Histone deacetylase inhibitors in Pediatric Brain Cancers: Biological Activities and therapeutic potential. Front Cell Dev Biol. 2020;8:1–14. https://doi.org/10.3389/fcell.2020.00546.
Taspinar Çakir, Güven, Denizler K, Rüstemoğlu. “The Effect of Trichostatin A on Radiosensitivity in Glioblastoma,” Proceedings, vol. 40, no. 1, p. 22, 2019, https://doi.org/10.3390/proceedings2019040022.
Savran B, Yerlikaya A, Erdoğan E, Genç O. “Anticancer Agent Ukrain and Bortezomib Combination is Synergistic in 4T1 Breast Cancer Cells,” Anticancer. Agents Med. Chem, vol. 14, no. 3, pp. 466–472, Feb. 2014, https://doi.org/10.2174/18715206113139990318.
Chou TC. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev. 2006;58(3):621–81. https://doi.org/10.1124/pr.58.3.10.
Lan F, Yang Y, Han J, Wu Q, Yu H, Yue X. Sulforaphane reverses chemo-resistance to temozolomide in glioblastoma cells by NF-κB-dependent pathway downregulating MGMT expression. Int J Oncol. 2016;48(2):559–68. https://doi.org/10.3892/ijo.2015.3271.
Staberg M, Michaelsen SR, Rasmussen RD, Villingshøj M, Poulsen HS, Hamerlik P. Inhibition of histone deacetylases sensitizes glioblastoma cells to lomustine. Cell Oncol. 2017;40(1):21–32. https://doi.org/10.1007/s13402-016-0301-9.
Kato Y, et al. Contributing factors of temozolomide resistance in MCF-7 tumor xenograft models. Cancer Biol Ther. 2007;6(6):891–7. https://doi.org/10.4161/cbt.6.6.4096.
Kitange GJ, et al. Induction of MGMT expression is associated with temozolomide resistance in glioblastoma xenografts. Neuro Oncol. 2009;11(3):281–91. https://doi.org/10.1215/15228517-2008-090.
Perazzoli G, et al. Temozolomide resistance in glioblastoma cell lines: implication of MGMT, MMR, P-glycoprotein and CD133 expression. PLoS ONE. 2015;10(10):1–23. https://doi.org/10.1371/journal.pone.0140131.
Meng CF, Zhu XJ, Peng G, Dai DQ. Role of histone modifications and DNA methylation in the regulation of O6-methylguanine-DNA methyltransferase gene expression in human stomach cancer cells. Cancer Invest. 2010;28(4):331–9. https://doi.org/10.3109/07357900903179633.
Danam RP, Howell SR, Brent TP, Harris LC. Epigenetic regulation of O6-methylguanine-DNA methyltransferase gene expression by histone acetylation and methyl-CpG binding proteins. Mol Cancer Ther. 2005;4(1):61–9. https://doi.org/10.1158/1535-7220.127.116.11.
Bhakat KK, Mitra S. Regulation of the human O6-methylguanine-DNA methyltransferase gene by transcriptional coactivators cAMP response element-binding protein-binding protein and p300. J Biol Chem. 2000;275(44):34197–204. https://doi.org/10.1074/jbc.M005447200.
Shinsato Y, et al. Reduction of MLH1 and PMS2 confers temozolomide resistance and is associated with recurrence of glioblastoma. Oncotarget. 2013;4(12):2261–70. https://doi.org/10.18632/oncotarget.1302.
Meng CF, Su B, Li W. DNA demethylation is superior to histone acetylation for reactivating cancer-associated genes in ovarian cancer cells. Mol Med Rep. 2011;4(6):1273–8. https://doi.org/10.3892/mmr.2011.557.
Imesch P, Dedes KJ, Furlato M, Fink D, Fedier A. MLH1 protects from resistance acquisition by the histone deacetylase inhibitor trichostatin A in colon tumor cells. Int J Oncol. 2009;35(3):631–40. https://doi.org/10.3892/ijo_00000375.
Kondo Y, Shen L, Issa J-PJ. Critical role of histone methylation in tumor suppressor gene silencing in Colorectal Cancer. Mol Cell Biol. 2003;23(1):206–15. https://doi.org/10.1128/mcb.23.1.206-215.2003.
McFaline-Figueroa JL, et al. Minor changes in expression of the mismatch repair protein MSH2 exert a major impact on glioblastoma response to temozolomide. Cancer Res. 2015;75:3127–38. https://doi.org/10.1158/0008-5472.CAN-14-3616.
Castro GN, et al. Effects of temozolomide (TMZ) on the expression and interaction of heat shock proteins (HSPs) and DNA repair proteins in human malignant glioma cells. Cell Stress Chaperones. 2015;20(2):253–65. https://doi.org/10.1007/s12192-014-0537-0.
Ponnusamy L, Mahalingaiah PKS, Chang YW, Singh KP. “Reversal of epigenetic aberrations associated with the acquisition of doxorubicin resistance restores drug sensitivity in breast cancer cells,” Eur. J. Pharm. Sci, vol. 123, no. June, pp. 56–69, 2018, https://doi.org/10.1016/j.ejps.2018.07.028.
This study was funded by The Scientific and Technological Research Council of Turkey (Grant No: 1919B011802106) and the Van Yuzuncu Yıl University Scientific Research Council. Author M.G received the grants.
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Güven, M., Taşpınar, F., Denizler-Ebiri, F.N. et al. The antagonistic effects of temozolomide and trichostatin a combination on MGMT and DNA mismatch repair pathways in Glioblastoma. Med Oncol 40, 223 (2023). https://doi.org/10.1007/s12032-023-02079-6