Spontaneous in vitro senescence of glioma cells confirmed by an antibody against IDH1R132H

Anticancer Res. 2014 Jun;34(6):2859-67.

Abstract

Background: We have recently suggested that glioblastoma cells become spontaneously senescent in cell culture conditions. The antibody specific against IDH1(R132H) offers the perfect opportunity to verify this hypothesis.

Materials and methods: We analyzed the features of senescence in 8 glioma cell cultures showing the IDH1(R132H) mutation based on combination of immunocytochemistry, enzymo-cytochemistry, BrdU incorporation assay and real-time microscopic observation.

Results: We report that glioma cells showing the IDH1(R132H) mutation become rapidly and spontaneously senescent in vitro. Senescence was observed in both classical and novel serum-free cell culture conditions. Importantly, the senescent IDH1(R132H)-positive cells showed the expression of stemness marker (SOX2).

Conclusion: In vitro senescence appeared to be the main reason of the difficulties in any kind culturing of glioma cells. 3D cell cultures prolonged the survival and in vitro proliferation of neoplastic IDH1(R132H)-positive cells, however, did not enhance the stabilization efficiency. Senescence of glioma cells is spontaneously triggered in vitro, which offers the opportunity of potential new therapeutic strategies based on this phenomenon.

Keywords: 3D cell culture; IDH1 gene; astrocytoma; glioblastoma; glioma; senescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal*
  • Blotting, Western
  • Brain Neoplasms / genetics
  • Brain Neoplasms / immunology
  • Brain Neoplasms / pathology*
  • Cell Culture Techniques
  • Cell Movement
  • Cell Proliferation
  • Cellular Senescence / physiology*
  • Flow Cytometry
  • Glioma / genetics
  • Glioma / immunology
  • Glioma / pathology*
  • Humans
  • Isocitrate Dehydrogenase / genetics*
  • Isocitrate Dehydrogenase / immunology
  • Isocitrate Dehydrogenase / metabolism
  • Mutation / genetics*
  • Neoplastic Stem Cells / metabolism
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • SOXB1 Transcription Factors / metabolism
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • RNA, Messenger
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • Isocitrate Dehydrogenase
  • IDH1 protein, human