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Tumor Biology

, Volume 35, Issue 5, pp 4479–4488 | Cite as

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

  • 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
Research Article

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.

Keywords

High-grade glioma PTPN6 SHP1 Survival Chemotherapy Methylation 

Notes

Acknowledgments

The authors would like to thank Prof. Frank Böhmer, Jena University, for kindly providing the pRK5PTPN6 vector; Dr. H Hedman, Umea University, the UCSF Tissue Bank, and Dr. JS Guillamo for providing us with the glioma cell lines used in the experiments; and the Science for Life Laboratory BioVis Technology Platform in Uppsala for supporting the flow cytometry analyses. We would also like to express our gratitude for the financial support from the Cancer Foundation at Gavle Hospital, the Research Fund at the Department of Oncology, Uppsala University Hospital, the Swedish Cancer Society, the Swedish Research Council, and the Knut and Alice Wallenberg Foundation.

Conflicts of interest

None

Supplementary material

13277_2013_1590_Fig6_ESM.jpg (41 kb)
Online resource 1

Mean dose response curves from FMCA analysis for MGMT and PTPN6 overexpressing clones and control clones. PTPN6 clones (n = 5) were less sensitive than control clones (n = 5) to the drugs bortezomib (A), cisplatin (B) and melphalan (C). (JPEG 41 kb)

13277_2013_1590_MOESM1_ESM.tif (2.3 mb)
High resolution image (TIFF 2334 kb)
13277_2013_1590_Fig7_ESM.jpg (22 kb)
Online resource 2

Mean cell cycle distribution of three PTPN6 and three control clones with or without bortezomib treatment. Bortezomib increased the percentage of cells in G0/G1 phase (p < 0.05) and decreased the percentage of cells in S and G2/M phases (p < 0.05) after 72 h in both PTPN6 overexpressing clones and control clones. (JPEG 22 kb)

13277_2013_1590_MOESM2_ESM.tif (205 kb)
High resolution image (TIFF 205 kb)

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Linda Sooman
    • 1
  • Simon Ekman
    • 1
  • Georgios Tsakonas
    • 1
  • Archita Jaiswal
    • 2
  • Sanjay Navani
    • 2
  • Per-Henrik Edqvist
    • 3
    • 4
  • Fredrik Pontén
    • 3
    • 4
  • Stefan Bergström
    • 1
  • Mikael Johansson
    • 5
  • Xuping Wu
    • 1
  • Erik Blomquist
    • 1
  • Michael Bergqvist
    • 1
  • Joachim Gullbo
    • 1
    • 6
  • Johan Lennartsson
    • 7
  1. 1.Rudbeck Laboratory, Section of Oncology, Department of Radiology, Oncology and Radiation SciencesUppsala UniversityUppsalaSweden
  2. 2.The Human Protein Atlas ProjectMumbaiIndia
  3. 3.Rudbeck Laboratory, Department of Immunology, Genetics and PathologyUppsala UniversityUppsalaSweden
  4. 4.Rudbeck Laboratory Department of Immunology, Genetics and Pathology and Science for Life LaboratoryUppsala UniversityUppsalaSweden
  5. 5.Oncology, Department of Radiation SciencesUmeå University HospitalUmeåSweden
  6. 6.Section of Clinical Pharmacology, Department of Medical SciencesUppsala University HospitalUppsalaSweden
  7. 7.Ludwig Institute for Cancer ResearchUppsala UniversityUppsalaSweden

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