Effects of Some Growth Factors and Cytokines on the Expression of the Repair Enzyme MGMT and Protein MARP in Human Cells In Vitro
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The inducible repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) eliminates O6-methylguanine adducts in DNA and protects the cells from damaging effects of alkylating agents. We have found that anti-MGMT antibodies recognize both the MGMT protein with a mol. weight ~ 24 kDa and a protein with a mol. weight ~ 48 kDa, which was named MARP (anti-methyltransferase antibody recognizable protein). A number of growth factors and cytokines were shown to regulate the expression of MGMT and MARP proteins. The ranges of concentrations of several growth factors and cytokines that caused increasing or decreasing protein amounts in human cell cultures were determined. The results of special biological experiments have allowed us to assume a possible role of MARP in the repair of alkyl adducts in human cells.
KeywordsCytokine DNA damage Repair enzyme MGMT Growth factor Antibody Human cells in vitro
We are very grateful to Professors Korneluyk A.I., Kuchuk M.V., Drs. Chernykh S.I., Potopalsky A.I., and Luchakovskaya Yu.S. for providing the preparations of cytokines and alkylating agent.
The study was supported by the National Academy of Sciences of Ukraine (No. 0115U000355) and the Czech Research Infrastructure for Systems Biology C4SYS (Project No. LM2015055).
- Christmann M, Verbeek B, Roos WP, Kaina B (2011) O6-methylguanine-DNA methyltransferase (MGMT) in normal tissues and tumors: enzyme activity, promoter methylation and immunohistochemistry. Biochim Biophys Acta Rev Cancer 1816:179–190. https://doi.org/10.1016/j.bbcan.2011.06.002 CrossRefGoogle Scholar
- Ishiguro K, Shyam K, Penketh PG, Baumann RP, Sartorelli AC, Rutherford TJ, Ratner ES (2013) Expression of O6-methylguanine-DNA methyltransferase examined by alkyl-transfer assays, methylation-specific PCR and western blots in tumors and matched normal tissue. J Cancer Ther 4:919–931CrossRefPubMedPubMedCentralGoogle Scholar
- Jiang XB, Hu B, He DS, Mao ZG, Wang X, Song BB, Zhu YH, Wang HJ (2015) Expression profiling of O6-methylguanine-DNA-methyltransferase in prolactinomas: a correlative study of promoter methylation and pathological features in 136 cases. BMC Cancer. https://doi.org/10.1186/s12885-015-1595-0 Google Scholar
- Kohsaka S, Tanak S (2013) Chemotherapeutic agent for glioma. In: Clinical management and evolving novel therapeutic strategies for patients with brain tumors, vol 19. InTech, p 415–438. https://doi.org/10.5772/45956
- Latypov VF, Tubbs JL, Watson AJ, Marriott AS, McGown G, Thorn-croft M, Wilkinson OJ, Senthong P, Butt A, Arvai AS, Millington CL, Povey AC, Williams DM, Santibanez-Koref MF, Tainer JA, Margison GP (2012) Atl1regulates choice between global genome and transcription-coupled repair of O(6)-alkylguanines. Mol Cell 47:50–60CrossRefPubMedPubMedCentralGoogle Scholar
- Lukash LL, Boldt J, Pegg AE, Dolan ME, Maher VM, McCormic JJ (1991) Effect of O6-alkylguanine-DNA alkyltransferase on the frequency and spectrum of mutations induced by N-methyl-N′-nitro-N-nitrosoguanidine in the HPRT gene of diploid human fibroblasts. Mutat Res 250:397–409CrossRefPubMedGoogle Scholar
- Macewicz LL, Kushniruk VO, Iatsyshyna AP, Kotsarenko KV, Lylo VV, Akopyan GR, Huleuk NL, Mykytenko DO, Lukash LL (2013) Correlation of mutagenesis level with expression of reparative enzyme O6-methylguanine-DNA methyltransferase during establishment of cell lines in vitro. Biopolym Cell 29:480–486CrossRefGoogle Scholar
- Stepanenko A, Andreieva S, Korets K, Mykytenko D, Baklaushev V, Huleyuk N, Kovalova O, Kotsarenko K, Chekhonin V, Vassetzky Y, Avdieiev S, Dmitrenko V (2016) Temozolomide promotes genomic and phenotypic changes in glioblastoma cells. Cancer Cell Int. https://doi.org/10.1186/s12935-016-0311-8 PubMedPubMedCentralGoogle Scholar
- Wang Y, Li J, Tohti M, Hu Y, Wang S, Li W, Lu Z, Ma C (2014) The expression profile of Dopamine D2 receptor, MGMT and VEGF in different histological subtypes of pituitary adenomas: a study of 197 cases and indications for the medical therapy. J Exp Clin Cancer Res. https://doi.org/10.1186/s13046-014-0056-y Google Scholar
- Zheng M, Bocangel D, Ramesh R, Ekmekcioglu S, Poindexter N, Grimm EA, Chada S (2008) Interleukin-24 overcomes temozolomide resistance and enhances cell death by down-regulation of O6-methylguanine-DNA methyltransferase in human melanoma cells. Mol Cancer Ther 7:3842–3851CrossRefPubMedPubMedCentralGoogle Scholar