The influence of DNA repair enzyme MGMT and associated MARP protein on cytotoxic effect of alkylating compounds in human cell cultures

  • K. V. Kotsarenko Institute of Molecular Biology and Genetics, NAS of Ukraine, Ukraine, 03680, Kyiv, Akademika Zabolotnogo str., 150
  • O. A. Akulenko Taras Shevchenko National University of Kyiv, Ukraine, 01601, Kyiv, Volodymyrska str., 64/13
  • V. V. Lylo Institute of Molecular Biology and Genetics, NAS of Ukraine, Ukraine, 03680, Kyiv, Akademika Zabolotnogo str., 150
  • T. P. Ruban Institute of Molecular Biology and Genetics, NAS of Ukraine, Ukraine, 03680, Kyiv, Akademika Zabolotnogo str., 150
  • L. L. Macewicz Institute of Molecular Biology and Genetics, NAS of Ukraine, Ukraine, 03680, Kyiv, Akademika Zabolotnogo str., 150
  • A. Ya. Glavatskiy SI “Acad. Romodanov Institute of Neurosurgery, NAMS of Ukraine”, Ukraine, 04050, Kyiv, P. Majborody str., 32
  • O. V. Markova SI “Acad. Romodanov Institute of Neurosurgery, NAMS of Ukraine”, Ukraine, 04050, Kyiv, P. Majborody str., 32
  • K. A. Kardash SI “Acad. Romodanov Institute of Neurosurgery, NAMS of Ukraine”, Ukraine, 04050, Kyiv, P. Majborody str., 32
  • I. M. Shuba SI “Acad. Romodanov Institute of Neurosurgery, NAMS of Ukraine”, Ukraine, 04050, Kyiv, P. Majborody str., 32
  • L. L. Lukash Institute of Molecular Biology and Genetics, NAS of Ukraine, Ukraine, 03680, Kyiv, Akademika Zabolotnogo str., 150

Abstract

Aim. To compare the effect of various alkylating compounds on survival of human cells containing the MARP protein only or both MGMT and MARP proteins. Methods. The following cell lines were used in this study: Hep-2 (laryngeal carcinoma cells) and 4BL (adult stem cells). The cells were treated with nitrosoguanidine and commercial drugs Temodal and Amitozyn.  Cytotoxic effect of the drugs was assessed by cell colony formation assay. The presence of MGMT and MARP proteins was identified by Western-blot analysis using monoclonal anti-MGMT antibody (clone 23.2). Results. The dependence of sensitivity of Hep-2 cells containing both MGMT and MARP proteins to the low doses of alkylating compounds under study was nonlinear with a “plateau” stage. 4BL cells, containing only the MARP protein, were more sensitive to the action of alkylating compounds, especially of nitrosoguanidine. However, the treatment of 4BL cell line with relatively low concentrations of Temodal and Amitozyn drugs (up to 2 μM and up to 200μg/ml respectively) didn’t lead to a sharp decline in the number of viable cells and the concentration dependence was nonlinear, as in the case of Hep-2 cells. Conclusions. The dependence of Hep-2 and 4BL cells viability to the action of low concentrations of alkylating compounds Temodal and Amitozyn was nonlinear with a “plateau”. This suggests the fact that the repair of alkyl adducts caused by these drugs occurs in the cells, which involves not only reparative MGMT enzyme, but possibly MARP protein. However the presence of the MARP protein in the cells does not affect the repair of damages caused by the nitrosoguanidine.

Keywords: MGMT, MARP, reparation, alkylating compounds, cells viability.

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