Comparative study of autophagy inhibition by 3MA and CQ on Cytarabine-induced death of leukaemia cells

  • Caroline Palmeira-dos-Santos
  • Gustavo J. S. Pereira
  • Christiano M. V. Barbosa
  • Aron Jurkiewicz
  • Soraya S. Smaili
  • Claudia BincolettoEmail author
Original Article – Cancer Research



As the molecular mechanisms of Cytarabine, one of the most important drugs used in the leukaemia’s treatment, are only partially understood and the role of autophagy on leukaemia development and treatment is only recently being investigated, in this study, by using Chloroquine (CQ) and 3-methyladenine (3MA) as autophagy inhibitors, we aim to evaluate the contribution of an autophagic mechanism to Cytarabine (AraC)-induced death of HL60 leukaemia cells.


Trypan blue exclusion and AnnexinV/PI assays were used to evaluate HL60 cell death under AraC treatment in the presence or absence of 3MA and CQ. Western blotting and immunofluorescence experiments were performed to show the involvement of apoptosis and autophagy protein expressions. Phenotypic characterization of HL60-treated cells was performed by using immunophenotyping. Clonogenic assays were applied to analyse clonal function of HL60-treated cells.


We observed that although autophagy inhibition by 3MA, but not CQ, increased the death of HL60 AraC cells after 24 h of treatment, no significant differences between AraC and AraC + 3MA-treated groups were observed by using clonogenic assay. In addition, increased number of immature (CD34+/CD38Lin−/low) HL60 cells was found in AraC and AraC-3MA groups when compared with control untreated cells.


Although AraC anti-leukaemia effects could be potentiated by 3MA autophagy inhibition after 24 h of exposure, leukaemia cell resistance, the main causes of treatment failure, is also promoted by autophagy initial stage impairment by 3MA, denoting the complex role of autophagy in leukaemia cells’ response to chemotherapy.


HL60 AraC 3-Methyladenine Chloroquine Autophagy Cell death Differentiation 



This study was supported by grants from Fundação do Amparo a Pesquisa do Estado de São Paulo (FAPESP—12/51215-4) and Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior (CAPES) fellowship. The authors would like to thank Hanako Hirata, Maria de Lourdes L. dos Santos, Bruno Palmieri and Cıcero R. dos Santos for their technical support.

Conflict of interest

The authors have declared no conflict of interest.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Caroline Palmeira-dos-Santos
    • 1
  • Gustavo J. S. Pereira
    • 1
  • Christiano M. V. Barbosa
    • 2
  • Aron Jurkiewicz
    • 1
  • Soraya S. Smaili
    • 1
  • Claudia Bincoletto
    • 1
    Email author
  1. 1.Departamento de Farmacologia, Escola Paulista de Medicina (EPM)Universidade Federal de São Paulo (UNIFESP)São PauloBrazil
  2. 2.Departamento de Bioquímica, Escola Paulista de Medicina (EPM)Universidade Federal de São Paulo (UNIFESP)São PauloBrazil

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