Endogenous GD3 ganglioside induces apoptosis in U-1242 MG glioma cells

H. E. Saqr, O. Omran, S. Dasgupta, Robert K Yu, J. L. Oblinger, A. J. Yates

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

GD3 ganglioside induces apoptosis in several cell types, but the molecular events through which this occurs are largely unknown. We investigated the apoptotic effects of GD3 expression using U-1242 MG glioblastoma cells, as these cells synthesize almost exclusively GM3 and GM2 but not GD3. To express GD3 under the control of the TetOn system with minimum leakage, we modified the system by constructing a single tri-cistronic retrovirus vector containing three genes separated by two internal ribosome entry sites: (a) transcriptional silencer, tTS; (b) mutant of reverse transcriptional activator, rtTA2(S)-M2 (provided by H. Bujard, Heidelberg, Germany); and (c) enhanced green fluorescent protein (EGFP), as an indicator of the tri-cistronic gene expression. Using flow cytometry, we selected glioma cells (U1242MG-GD3 clone) that express high levels of GD3 in response to doxycycline. Expression of GD3 was associated with apoptosis as verified by annexin-V binding, TdT-mediated dUTPnick end-labelling assay (TUNEL)q2, and EGFP degradation. GD3-induced apoptosis occurred via caspase-8 activation, as GD3 caused cleavage of caspase-8 and inhibition of caspase-8 activation by zlETD-fmk minimized GD3-induced apoptosis.

Original languageEnglish (US)
Pages (from-to)1301-1314
Number of pages14
JournalJournal of Neurochemistry
Volume96
Issue number5
DOIs
StatePublished - Mar 1 2006

Fingerprint

Glioma
Caspase 8
Apoptosis
Chemical activation
Flow cytometry
Doxycycline
Annexin A5
Retroviridae
Glioblastoma
Gene expression
Labeling
Proteolysis
Germany
Assays
Flow Cytometry
Clone Cells
Genes
Gene Expression
Degradation
GD3 ganglioside

Keywords

  • Apoptosis
  • GD3
  • Ganglioside
  • Glioma

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Endogenous GD3 ganglioside induces apoptosis in U-1242 MG glioma cells. / Saqr, H. E.; Omran, O.; Dasgupta, S.; Yu, Robert K; Oblinger, J. L.; Yates, A. J.

In: Journal of Neurochemistry, Vol. 96, No. 5, 01.03.2006, p. 1301-1314.

Research output: Contribution to journalArticle

Saqr, HE, Omran, O, Dasgupta, S, Yu, RK, Oblinger, JL & Yates, AJ 2006, 'Endogenous GD3 ganglioside induces apoptosis in U-1242 MG glioma cells', Journal of Neurochemistry, vol. 96, no. 5, pp. 1301-1314. https://doi.org/10.1111/j.1471-4159.2005.03640.x
Saqr, H. E. ; Omran, O. ; Dasgupta, S. ; Yu, Robert K ; Oblinger, J. L. ; Yates, A. J. / Endogenous GD3 ganglioside induces apoptosis in U-1242 MG glioma cells. In: Journal of Neurochemistry. 2006 ; Vol. 96, No. 5. pp. 1301-1314.
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