Molecular Neurobiology

, Volume 55, Issue 5, pp 4185–4194 | Cite as

Low Dose of Doxorubicin Potentiates the Effect of Temozolomide in Glioblastoma Cells

  • Emilly Schlee Villodre
  • Franciele Cristina Kipper
  • Andrew Oliveira Silva
  • Guido Lenz
  • Patrícia Luciana da Costa Lopez


Glioblastoma (GBM) is an aggressive brain tumor with temozolomide (TMZ)-based chemotherapy as the main therapeutic strategy. Doxorubicin (DOX) is not used in gliomas due to its low bioavailability in the brain; however, new delivery strategies and low doses may be effective in the long term, especially as part of a drug cocktail. Our aim was to evaluate the chronic effects of low doses of DOX and TMZ in GBM. Human U87-ATCC cells and a primary GBM culture were chronically treated with TMZ (5 μM) and DOX (1 and 10 nM) alone or combined. DOX resulted in a reduction in the number of cells over a period of 35 days and delayed the cell regrowth. In addition, DOX induced cell senescence and reduced tumor sphere formation and the proportion of NANOG- and OCT4-positive cells after 7 days. Low doses of TMZ potentiated the effects of DOX on senescence and sphere formation. This combined response using low doses of DOX may pave the way for its use in glioma therapy, with new technologies to overcome its low blood–brain barrier permeability.


Glioblastoma Temozolomide Doxorubicin Cell senescence Cell proliferation Cancer stem cell 



This work was supported by CAPES Probitec 004/2012 and FAPERGS Pronem 11/2072-2. ESV and PLCL received CAPES fellowships and GL receives CNPq fellowship.

Compliance with Ethical Standards

The ethical committee at the UFRGS (n. 420.856) and PUCRS (n. 429.849) approved the use of LS12 cells.

Supplementary material

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Fig. S1

Flow cytometry analysis. (JPEG 72 kb)

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High-resolution image. (EPS 1112 kb)
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Fig. S2

Cumulative population doubling analysis of the primary GBM cell, LS12. (JPEG 252 kb)

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High-resolution image. (EPS 2072 kb)
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Fig. S3

Apoptosis and necrosis analysis after treatment with DOX and TMZ after 24 h. (JPEG 46 kb)

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High-resolution image. (EPS 1164 kb)
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Fig. S4

Nuclear morphometric analysis (NMA) after DOX 1 and 10 nM and TMZ 5 μM treatment. (JPEG 194 kb)

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High-resolution image. (EPS 28841 kb)
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Fig. S5

Number of spheres and protein quantification after 7 days of treatment. (JPEG 205 kb)

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High-resolution image. (EPS 1343 kb)
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Fig. S6

Radar graph was used in order to integrate all the results analysis using different combinations. (JPEG 262 kb)

12035_2017_611_MOESM6_ESM.eps (2.2 mb)
High-resolution image. (EPS 2223 kb)


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Emilly Schlee Villodre
    • 1
  • Franciele Cristina Kipper
    • 1
  • Andrew Oliveira Silva
    • 1
  • Guido Lenz
    • 1
    • 2
    • 3
  • Patrícia Luciana da Costa Lopez
    • 1
    • 4
    • 5
    • 6
  1. 1.Laboratory of Cellular Plasticity and Signaling, Department of Biophysics, Institute of BiosciencesFederal University of Rio Grande do Sul, UFRGSPorto AlegreBrazil
  2. 2.Center of BiotechnologyFederal University of Rio Grande do Sul, UFRGSPorto AlegreBrazil
  3. 3.Departamento de Biofísica, Instituto de BiociênciasUniversidade Federal do Rio Grande do Sul (UFRGS)Porto AlegreBrazil
  4. 4.Graduate Program in Gastroenterology and Hepatology, School of MedicineFederal University of Rio Grande do Sul, UFRGSPorto AlegreBrazil
  5. 5.Research CenterHospital de Clínicas de Porto Alegre, HCPAPorto AlegreBrazil
  6. 6.Centro de Pesquisas ExperimentalHospital de Clínicas de Porto Alegre (HCPA)Porto AlegreBrazil

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