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01-05-2014 | Research Article

PTPN6 expression is epigenetically regulated and influences survival and response to chemotherapy in high-grade gliomas

Authors: Linda Sooman, Simon Ekman, Georgios Tsakonas, Archita Jaiswal, Sanjay Navani, Per-Henrik Edqvist, Fredrik Pontén, Stefan Bergström, Mikael Johansson, Xuping Wu, Erik Blomquist, Michael Bergqvist, Joachim Gullbo, Johan Lennartsson

Published in: Tumor Biology | Issue 5/2014

Abstract

The prognosis of high-grade glioma patients is poor, and the tumors are characterized by resistance to therapy. The aims of this study were to analyze the prognostic value of the expression of the protein tyrosine phosphatase non-receptor type 6 (PTPN6, also referred to as SHP1) in high-grade glioma patients, the epigenetic regulation of the expression of PTPN6, and the role of its expression in chemotherapy resistance in glioma-derived cells. PTPN6 expression was analyzed with immunohistochemistry in 89 high-grade glioma patients. Correlation between PTPN6 expression and overall survival was analyzed with Kaplan-Meier univariate analysis and Cox regression multivariate analysis. Differences in drug sensitivity to a panel of 16 chemotherapeutic drugs between PTPN6-overexpressing clones and control clones were analyzed in vitro with the fluorometric microculture cytotoxicity assay. Cell cycle analysis was done with Krishan staining and flow cytometry. Apoptosis was analyzed with a cell death detection ELISA kit as well as cleaved caspase-3 and caspase-9 Western blotting. Autophagy was analyzed with LC3B Western blotting. Methylation of the PTPN6 promoter was analyzed with bisulfite pyrosequencing, and demethylation of PTPN6 was done with decitabine treatment. The PTPN6 expression correlated in univariate analysis to poor survival for anaplastic glioma patients (p = 0.026). In glioma-derived cell lines, overexpression of PTPN6 caused increase resistance (p < 0.05) to the chemotherapeutic drugs bortezomib, cisplatin, and melphalan. PTPN6 expression did not affect bortezomib-induced cell cycle arrest, apoptosis, or autophagy. Low PTPN6 promoter methylation correlated to protein expression, and the protein expression was increased upon demethylation in glioma-derived cells. PTPN6 expression may be a factor contributing to poor survival for anaplastic glioma patients, and in glioma-derived cells, its expression is epigenetically regulated and influences the response to chemotherapy.

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Title: PTPN6 expression is epigenetically regulated and influences survival and response to chemotherapy in high-grade gliomas
Authors: Linda Sooman
Simon Ekman
Georgios Tsakonas
Archita Jaiswal
Sanjay Navani
Per-Henrik Edqvist
Fredrik Pontén
Stefan Bergström
Mikael Johansson
Xuping Wu
Erik Blomquist
Michael Bergqvist
Joachim Gullbo
Johan Lennartsson
Publication date: 01-05-2014
Publisher: Springer Netherlands
Published in: Tumor Biology / Issue 5/2014
Print ISSN: 1010-4283
Electronic ISSN: 1423-0380
DOI: https://doi.org/10.1007/s13277-013-1590-5

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