Open Access

The DNA damage/repair cascade in glioblastoma cell lines after chemotherapeutic agent treatment

  • Authors:
    • Laura Annovazzi
    • Valentina Caldera
    • Marta Mellai
    • Chiara Riganti
    • Luigi Battaglia
    • Daniela Chirio
    • Antonio Melcarne
    • Davide Schiffer
  • View Affiliations

  • Published online on: April 16, 2015     https://doi.org/10.3892/ijo.2015.2963
  • Pages: 2299-2308
  • Copyright: © Annovazzi et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Therapeutic resistance in glioblastoma multiforme (GBM) has been linked to a subpopulation of cells with stem cell-like properties, the glioma stem cells (GSCs), responsible for cancer progression and recurrence. This study investigated the in vitro cytotoxicity of three chemotherapeutics, temozolomide (TMZ), doxorubicin (Dox) and paclitaxel (PTX) on glioma cell lines, by analyzing the molecular mechanisms leading to DNA repair and cell resistance, or to cell death. The drugs were tested on 16 GBM cell lines, grown as neurospheres (NS) or adherent cells (AC), by studying DNA damage occurrence by Comet assay, the expression by immunofluorescence and western blotting of checkpoint/repair molecules and apoptosis. The three drugs were able to provoke a genotoxic injury and to inhibit dose- and time-dependently cell proliferation, more evidently in AC than in NS. The first cell response to DNA damage was the activation of the damage sensors (p-ATM, p-53BP1, γ-H2AX), followed by repair effectors; the expression of checkpoint/repair molecules appeared higher in NS than in AC. The non-homologous repair pathway (NHEJ) seemed more involved than the homologous one (HR). Apoptosis occurred after long treatment times, but only a small percentage of cells in NS underwent death, even at high drug concentration, whereas most cells survived in a quiescent state and resumed proliferation after drug removal. In tumor specimens, checkpoint/repair proteins were constitutively expressed in GBMs, but not in low-grade gliomas.
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June-2015
Volume 46 Issue 6

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Annovazzi L, Caldera V, Mellai M, Riganti C, Battaglia L, Chirio D, Melcarne A and Schiffer D: The DNA damage/repair cascade in glioblastoma cell lines after chemotherapeutic agent treatment. Int J Oncol 46: 2299-2308, 2015.
APA
Annovazzi, L., Caldera, V., Mellai, M., Riganti, C., Battaglia, L., Chirio, D. ... Schiffer, D. (2015). The DNA damage/repair cascade in glioblastoma cell lines after chemotherapeutic agent treatment. International Journal of Oncology, 46, 2299-2308. https://doi.org/10.3892/ijo.2015.2963
MLA
Annovazzi, L., Caldera, V., Mellai, M., Riganti, C., Battaglia, L., Chirio, D., Melcarne, A., Schiffer, D."The DNA damage/repair cascade in glioblastoma cell lines after chemotherapeutic agent treatment". International Journal of Oncology 46.6 (2015): 2299-2308.
Chicago
Annovazzi, L., Caldera, V., Mellai, M., Riganti, C., Battaglia, L., Chirio, D., Melcarne, A., Schiffer, D."The DNA damage/repair cascade in glioblastoma cell lines after chemotherapeutic agent treatment". International Journal of Oncology 46, no. 6 (2015): 2299-2308. https://doi.org/10.3892/ijo.2015.2963