Epigenetic regulation of the secreted frizzled-related protein family in human glioblastoma multiforme

L. Schiefer, M. Visweswaran, V. Perumal, F. Arfuso, D. Groth, P. Newsholme, S. Warrier, A. Dharmarajan

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Glioblastoma multiforme (GBM) are intracranial tumors of the central nervous system and the most lethal among solid tumors. Current therapy is palliative and is limited to surgical resection followed by radiation therapy and temozolomide treatment. Aberrant WNT pathway activation mediates not only cancer cell proliferation but also promotes radiation and chemotherapeutic resistance. WNT antagonists such as the secreted frizzled-related protein (sFRP) family have an ability to sensitize glioma cells to chemotherapeutics, decrease proliferation rate and induce apoptosis. During tumor development, sFRP genes (1-5) are frequently hypermethylated, causing transcriptional silencing. We investigated a possible involvement of methylation-mediated silencing of the sFRP gene family in human GBM using four human glioblastoma cell lines (U87, U138, A172 and LN18). To induce demethylation of the DNA, we inhibited DNA methyltransferases through treatment with 5-azacytidine. Genomic DNA, RNA and total protein were isolated from GBM cells before and after treatment. We utilized bisulfite modification of genomic DNA to examine the methylation status of the respective sFRP promoter regions. Pharmacological demethylation of the GBM cell lines demonstrated a loss of methylation in sFRP promoter regions, as well as an increase in sFRP gene-specific mRNA abundance. Western blot analysis demonstrated an increased protein expression of sFRP-4 and increased levels of phosphorylated-β-catenin. These data indicate an important role of methylation-induced gene silencing of the sFRP gene family in human GBM.

Original languageEnglish
Pages (from-to)297-303
Number of pages7
JournalCancer Gene Therapy
Volume21
Issue number7
DOIs
Publication statusPublished - 2014

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Glioblastoma
Epigenomics
Methylation
DNA
temozolomide
Genetic Promoter Regions
Genes
Azacitidine
Cell Line
Catenins
Central Nervous System Neoplasms
Neoplasms
Methyltransferases
Gene Silencing
Palliative Care
Glioma
FRZB protein
Proteins
Radiotherapy
Therapeutics

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Cancer Research

Cite this

Schiefer, L., Visweswaran, M., Perumal, V., Arfuso, F., Groth, D., Newsholme, P., ... Dharmarajan, A. (2014). Epigenetic regulation of the secreted frizzled-related protein family in human glioblastoma multiforme. Cancer Gene Therapy, 21(7), 297-303. https://doi.org/10.1038/cgt.2014.30
Schiefer, L. ; Visweswaran, M. ; Perumal, V. ; Arfuso, F. ; Groth, D. ; Newsholme, P. ; Warrier, S. ; Dharmarajan, A. / Epigenetic regulation of the secreted frizzled-related protein family in human glioblastoma multiforme. In: Cancer Gene Therapy. 2014 ; Vol. 21, No. 7. pp. 297-303.
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Schiefer, L, Visweswaran, M, Perumal, V, Arfuso, F, Groth, D, Newsholme, P, Warrier, S & Dharmarajan, A 2014, 'Epigenetic regulation of the secreted frizzled-related protein family in human glioblastoma multiforme', Cancer Gene Therapy, vol. 21, no. 7, pp. 297-303. https://doi.org/10.1038/cgt.2014.30

Epigenetic regulation of the secreted frizzled-related protein family in human glioblastoma multiforme. / Schiefer, L.; Visweswaran, M.; Perumal, V.; Arfuso, F.; Groth, D.; Newsholme, P.; Warrier, S.; Dharmarajan, A.

In: Cancer Gene Therapy, Vol. 21, No. 7, 2014, p. 297-303.

Research output: Contribution to journalArticle

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T1 - Epigenetic regulation of the secreted frizzled-related protein family in human glioblastoma multiforme

AU - Schiefer, L.

AU - Visweswaran, M.

AU - Perumal, V.

AU - Arfuso, F.

AU - Groth, D.

AU - Newsholme, P.

AU - Warrier, S.

AU - Dharmarajan, A.

PY - 2014

Y1 - 2014

N2 - Glioblastoma multiforme (GBM) are intracranial tumors of the central nervous system and the most lethal among solid tumors. Current therapy is palliative and is limited to surgical resection followed by radiation therapy and temozolomide treatment. Aberrant WNT pathway activation mediates not only cancer cell proliferation but also promotes radiation and chemotherapeutic resistance. WNT antagonists such as the secreted frizzled-related protein (sFRP) family have an ability to sensitize glioma cells to chemotherapeutics, decrease proliferation rate and induce apoptosis. During tumor development, sFRP genes (1-5) are frequently hypermethylated, causing transcriptional silencing. We investigated a possible involvement of methylation-mediated silencing of the sFRP gene family in human GBM using four human glioblastoma cell lines (U87, U138, A172 and LN18). To induce demethylation of the DNA, we inhibited DNA methyltransferases through treatment with 5-azacytidine. Genomic DNA, RNA and total protein were isolated from GBM cells before and after treatment. We utilized bisulfite modification of genomic DNA to examine the methylation status of the respective sFRP promoter regions. Pharmacological demethylation of the GBM cell lines demonstrated a loss of methylation in sFRP promoter regions, as well as an increase in sFRP gene-specific mRNA abundance. Western blot analysis demonstrated an increased protein expression of sFRP-4 and increased levels of phosphorylated-β-catenin. These data indicate an important role of methylation-induced gene silencing of the sFRP gene family in human GBM.

AB - Glioblastoma multiforme (GBM) are intracranial tumors of the central nervous system and the most lethal among solid tumors. Current therapy is palliative and is limited to surgical resection followed by radiation therapy and temozolomide treatment. Aberrant WNT pathway activation mediates not only cancer cell proliferation but also promotes radiation and chemotherapeutic resistance. WNT antagonists such as the secreted frizzled-related protein (sFRP) family have an ability to sensitize glioma cells to chemotherapeutics, decrease proliferation rate and induce apoptosis. During tumor development, sFRP genes (1-5) are frequently hypermethylated, causing transcriptional silencing. We investigated a possible involvement of methylation-mediated silencing of the sFRP gene family in human GBM using four human glioblastoma cell lines (U87, U138, A172 and LN18). To induce demethylation of the DNA, we inhibited DNA methyltransferases through treatment with 5-azacytidine. Genomic DNA, RNA and total protein were isolated from GBM cells before and after treatment. We utilized bisulfite modification of genomic DNA to examine the methylation status of the respective sFRP promoter regions. Pharmacological demethylation of the GBM cell lines demonstrated a loss of methylation in sFRP promoter regions, as well as an increase in sFRP gene-specific mRNA abundance. Western blot analysis demonstrated an increased protein expression of sFRP-4 and increased levels of phosphorylated-β-catenin. These data indicate an important role of methylation-induced gene silencing of the sFRP gene family in human GBM.

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