Biopolym. Cell. 2013; 29(6):480-486.
http://dx.doi.org/10.7124/bc.00083D
Genomics, Transcriptomics and Proteomics
Correlation of mutagenesis level with expression of reparative enzyme O6-methylguanine DNA methyltransferase during establishment of cell lines in vitro
1Macewicz L. L., 1Kushniruk V. O., 1Iatsyshyna A. P., 1Kotsarenko K. V., 1Lylo V. V., 2Akopyan G. R., 2Huleuk N. L., 3Mykytenko D. O., 1Lukash L. L.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Institute of Hereditary Pathology MAS of Ukraine
    31a, Lysenko Str., Lviv, Ukraine, 79008
  3. "Nadiya" Clinic
    19A, M. Krivonosa, Kyiv, Ukraine, 03037

Abstract

Long-term cultivation of cell lines inevitably leads to genetic and epigenetic changes. The aim of this work was a comparative analysis of karyotype and level of the expression of reparative enzyme O6-methylguanine-DNA- methyltransferase (MGMT) at different stages of establishment and stabilization of human cell line 4BL and cell line of mouse germ cells G1. Methods. The set of methods was used to research the dynamics of karyotypes changes: the differential staining of chromosomes, FISH-method and comparative genomic hybridization. The level of MGMT expression was analyzed by PCR reaction and Western blot analysis. Results. General trends of establishment of mouse and human cell lines were revealed: at the first stage, which is characterized by increased structural instability of the genome, an increase in the MGMT expression was revealed while at the second stage of stabilization – a decrease in the expression. Therefore, almost complete disappearance of MGMT protein in unmodified form (24 kDa) is observed. Conclusions. Statistically significant correlation between MGMT repair enzyme and mutations induction processes during mammalian cell adaptation and cell line establishment to in vitro was described.
Keywords: cell line, karyotypic evolution, O6-methylguanine-DNA-methyltransferase, genomic instability
Full text: (PDF, in English)

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