Oncotarget

Research Papers:

Panobinostat mediated cell death: a novel therapeutic approach for osteosarcoma

André Wirries, Samir Jabari, Esther P. Jansen, Silvia Roth, Elizabeth Figueroa-Juárez, Thaddeus T. Wissniowski, Daniel Neureiter, Eckhard Klieser, Philipp Lechler, Steffen Ruchholtz, Detlef K. Bartsch, Christoph K. Boese and Pietro Di Fazio _

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Oncotarget. 2018; 9:32997-33010. https://doi.org/10.18632/oncotarget.26038

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Abstract

André Wirries1,8, Samir Jabari2, Esther P. Jansen1, Silvia Roth3, Elizabeth Figueroa-Juárez3, Thaddeus T. Wissniowski4, Daniel Neureiter5,6, Eckhard Klieser5,6, Philipp Lechler1, Steffen Ruchholtz1, Detlef K. Bartsch3, Christoph K. Boese7 and Pietro Di Fazio3

1Center of Orthopaedics and Trauma Surgery, Philipps University of Marburg, Baldingerstrasse 35043 Marburg, Germany

2Institute of Anatomy I, University of Erlangen-Nuremberg, 91054 Erlangen, Germany

3Department of Visceral, Thoracic and Vascular Surgery, Philipps University of Marburg, Baldingerstrasse 35043 Marburg, Germany

4Department of Gastroenterology and Endocrinology, Philipps University of Marburg, Baldingerstrasse 35043 Marburg, Germany

5Institute of Pathology, Paracelsus Medical University/Salzburger Landeskliniken (SALK), 5020 Salzburg, Austria

6Salzburg Cancer Research Institute, 5020 Salzburg, Austria

7Department of Orthopaedic and Trauma Surgery, University Hospital of Cologne, 50937 Cologne, Germany

8Orthopaedic Clinics, Hessing Foundation, 86199 Augsburg, Germany

Correspondence to:

Pietro Di Fazio, email: difazio@med.uni-marburg.de

Keywords: osteosarcoma; endoplasmic reticulum stress; autophagy; histone deacetylase inhibitor; cell death

Received: August 25, 2017     Accepted: August 16, 2018     Published: August 31, 2018

ABSTRACT

Osteosarcoma is an aggressive cancer with a poor long term prognosis. Neo-adjuvant poly-chemotherapy followed by surgical resection remains the standard treatment, which is restricted by multi-drug resistance. If first-line therapy fails, disease control and patient survival rate drop dramatically. We aimed to identify alternative apoptotic mechanisms induced by the histone deacetylase inhibitor panobinostat in osteosarcoma cells.

Saos-2, MG63 and U2-OS osteosarcoma cell lines, the immortalized human osteoblast line hFOB and the mouse embryo osteoblasts (MC3T3-E1) were treated with panobinostat. Real time viability and FACS confirmed the cytotoxicity of panobinostat. Cell stress/death related factors were analysed by RT-qPCR and western blot. Cell morphology was assessed by electron microscopy.

10 nM panobinostat caused cell viability arrest and death in all osteosarcoma and osteoblast cells. P21 up-regulation was observed in osteosarcoma cells, while over-expression of p73 was restricted to Saos-2 (TP53−/−).

Survivin and Bcl-2 were suppressed by panobinostat. Endoplasmic reticulum (ER) stress markers BiP, CHOP, ATF4 and ATF6 were induced in osteosarcoma cells. The un-spliced Xbp was no further detectable after treatment.

Autophagy players Beclin1, Map1LC3B and UVRAG transcripts over-expressed after 6 hours. Protein levels of Beclin1, Map1LC3B and p62 were up-regulated at 72 hours. DRAM1 was stable. Electron micrographs revealed the fragmentation and the disappearance of the ER and the statistically significant increase of autophagosome vesiculation after treatment.

Panobinostat showed a synergistic suppression of survival and promotion of cell death in osteosarcoma cells. Panobinostat offers new perspectives for the treatment of osteosarcoma and other malignant bone tumours.


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