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Effect of lomeguatrib–temozolomide combination on MGMT promoter methylation and expression in primary glioblastoma tumor cells

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Tumor Biology

Abstract

Temozolomide (TMZ) is commonly used in the treatment of glioblastoma (GBM). The MGMT repair enzyme (O 6-methylguanine-DNA methyltransferase) is an important factor causing chemotherapeutic resistance. MGMT prevents the formation of toxic effects of alkyl adducts by removing them from the DNA. Therefore, MGMT inhibition is an interesting therapeutic approach to circumvent TMZ resistance. The aim of the study was to investigate the effect of the combination of lomeguatrib (an MGMT inactivator) with TMZ, on MGMT expression and methylation. Primary cell cultures were obtained from GBM tumor tissues. The sensitivity of primary GBM cell cultures and GBM cell lines to TMZ, and to the combination of TMZ and lomeguatrib, was determined by a cytotoxicity assay (MTT). MGMT and p53 expression, and MGMT methylation were investigated after drug application. In addition, the proportion of apoptotic cells and DNA fragmentation was analyzed. The combination of TMZ and lomeguatrib in primary GBM cell cultures and glioma cell lines decreased MGMT expression, increased p53 expression, and did not change MGMT methylation. Moreover, apoptosis was induced and DNA fragmentation was increased in cells. In addition, we also showed that lomeguatrib–TMZ combination did not have any effect on the cell cycle. Finally, we determined that the sensitivity of each primary GBM cells and glioma cell lines to the lomeguatrib–TMZ combination was different and significantly associated with the structure of MGMT methylation. Our study suggests that lomeguatrib can be used with TMZ for GBM treatment, although further clinical studies will be needed so as to determine the feasibility of this therapeutic approach.

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Acknowledgements

The authors are grateful to Laura Stokes for help with editing the manuscript.

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Authors and Affiliations

  1. Department of Medical Biology, Ankara University School of Medicine, Ankara, Turkey

    Mehmet Taspinar, Seda Ilgaz, Tulin Ozkan & Asuman Sunguroğlu

  2. Department of Neurosurgery, Ankara University School of Medicine, Ankara, Turkey

    Mevci Ozdemir & Hasan Caglar Ugur

  3. Department of Biostatistics, Ankara University School of Medicine, Ankara, Turkey

    Derya Oztuna

  4. Department of Basic Oncology, Institute of Oncology, Hacettepe University School of Medicine, Ankara, Turkey

    Hande Canpinar

  5. IdiPaz Research Unit, La Paz University Hospital, Madrid, Spain

    Juan A. Rey

  6. Brain Tumor Biology Unit, University of Navarra School of Sciences, Pamplona, Spain

    Javier S. Castresana

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  1. Mehmet Taspinar
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Taspinar, M., Ilgaz, S., Ozdemir, M. et al. Effect of lomeguatrib–temozolomide combination on MGMT promoter methylation and expression in primary glioblastoma tumor cells. Tumor Biol. 34, 1935–1947 (2013). https://doi.org/10.1007/s13277-013-0738-7

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