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Extra-cellular matrix suppresses expression of the apoptosis mediator Fas by epigenetic DNA methylation

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Abstract

The extracellular matrix (ECM) of bone consists mainly of collagen type I, which induces osteoblastic differentiation and prevents apoptosis. Fas induces apoptosis in cells improperly adhering to ECM. Recently, it was described that Fas expression is modulated by epigenetic DNA methylation. Mouse MC3T3-E1 pre-osteoblastic cells were cultured either on collagen coated or on uncoated culture dishes for control. mRNA was isolated and gene expression was analyzed by quantitative RT–PCR. Furthermore, we measured global and specific DNA methylation. Compared to controls, cells cultured on collagen-coated dishes increased the expression of Runx2 and OCN indicating differentiation of pre-osteoblastic cells. Additionally, collagen up-regulated cyclin-A2 and down-regulated Fas expression suggesting increased cell multiplication. Furthermore, the expression of Dnmt1 and Hells, key mediators of the DNA-methylation process, was increased. As a consequence, we demonstrate that global DNA methylation and specific methylation of the Fas promoter was higher in MC3T3-E1 cells cultured on collagen when compared to controls. Investigation of signal transduction pathways by mean of inhibitors suggests that focal adhesion kinase, MAP- and Jun-kinases and AP-1 are involved in this process. In summary, we demonstrate that ECM prevents activation of Fas by epigenetic DNA-methylation.

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Acknowledgments

This study was supported by the Fonds zur Foerderung der wissenschaftlichen Forschung (FWF; The Austrian Science Fund) Project P20646-B11, the WGKK (Social Health Insurance Vienna), and the AUVA (Austrian Social Insurance for Occupational Risks). The authors thank to Dr. E. Paschalis for critical review of the manuscript and valuable comments.

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Authors and Affiliations

  1. Ludwig Boltzmann Institute of Osteology at the Hanusch Hospital of WGKK and AUVA Trauma Center Meidling, 4th Medical Department, Hanusch Hospital, Heinrich Collin-Str. 30, 1140, Vienna, Austria

    Roman Thaler, Silvia Spitzer, Klaus Klaushofer & Franz Varga

  2. Ludwig Boltzmann Institute for Leukemia Research and Hematology, Hanusch Hospital, and Ludwig Boltzmann Cluster Oncology, Vienna, Austria

    Heidrun Karlic

Authors
  1. Roman Thaler
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  2. Heidrun Karlic
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  3. Silvia Spitzer
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  4. Klaus Klaushofer
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  5. Franz Varga
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Correspondence to Franz Varga.

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Thaler, R., Karlic, H., Spitzer, S. et al. Extra-cellular matrix suppresses expression of the apoptosis mediator Fas by epigenetic DNA methylation. Apoptosis 15, 728–737 (2010). https://doi.org/10.1007/s10495-010-0462-3

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