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The Scientific World Journal
Volume 2014, Article ID 450390, 9 pages
http://dx.doi.org/10.1155/2014/450390
Research Article

Synergistic Antitumor Effect of Doxorubicin and Tacrolimus (FK506) on Hepatocellular Carcinoma Cell Lines

1Istituto Nazionale per lo Studio E la Cura dei Tumori “Fondazione Giovanni Pascale”, IRCCS, 80131 Naples, Italy
2Biochemistry, Biophysics and General Pathology Department, Second University of Naples, 80138 Naples, Italy
3Department of Onco-Hematology, S.G. Moscati Hospital, 83100 Avellino, Italy

Received 5 November 2013; Accepted 24 December 2013; Published 20 February 2014

Academic Editors: C.-H. Chen and D. Morris

Copyright © 2014 Francesca Capone et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Hepatocellular carcinoma is the fifth most common cancer worldwide and shows a complex clinical course, poor response to pharmacological treatment, and a severe prognosis. Thus, the aim of this study was to investigate whether tacrolimus (FK506) has synergistic antitumor effects with doxorubicin on two human hepatocellular carcinoma cell lines, Huh7 and HepG2. Cell viability was analyzed by Sulforhodamine B assay and synergic effect was evaluated by the software CalcuSyn. Cell apoptosis was evaluated using Annexin V and Dead Cell assay. Apoptosis-related protein PARP-1 cleaved and autophagy-related protein expressions (Beclin-1 and LC3B) were measured by western blotting analysis. Cytokines concentration in cellular supernatants after treatments was studied by Bio-Plex assay. Interestingly the formulation with doxorubicin and tacrolimus induced higher cytotoxicity level on tumor cells than single treatment. Moreover, our results showed that the mechanisms involved were (i) a strong cell apoptosis induction, (ii) contemporaneous decrease of autophagy activation, understood as prosurvival process, and (iii) downregulation of proinflammatory cytokines. In conclusion, future studies could relate to the doxorubicin/tacrolimus combination effects in mice models bearing HCC in order to see if this formulation could be useful in HCC treatment.