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The role of glycogen synthase kinase-3β in glioma cell apoptosis induced by remifentanil

Abstract

The aim of malignant glioma treatment is to inhibit tumor cell proliferation and induce tumor cell apoptosis. Remifentanil is a clinical anesthetic drug that can activate the N-methyl-D-aspartate (NMDA) receptor. NMDA receptor signaling activates glycogen synthase kinase-3β (GSK-3β). Discovered some 32 years ago, GSK-3β was only recently considered as a therapeutic target in cancer treatment. The purpose of this study was to assess whether remifentanil can induce the apoptosis of C6 cells through GSK-3β activation. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was used to detect cell viability. Hoechst 33342 staining and flow cytometry were used to detect cell apoptosis. The effect of GSK-3β activation was detected using a GSK-3β activation assay kit and 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8), a potent and selective small molecule inhibitor of GSK-3β. The MTT assay indicated that remifentanil induced C6 cell death in a concentration- and time-dependent manner. Hoechst 33342 staining and flow cytometry showed that remifentanil significantly induced C6 cell apoptosis. The measurement of GSK-3β activation showed that remifentanil increased the cellular level of GSK-3β. All of these toxic effects can be attenuated by treatment with TDZD-8. These results suggest that remifentanil is able to induce C6 cell apoptosis through GSK-3β activation, which provides a basis for its potential use in the treatment of malignant gliomas.

Abbreviations

FBS:

fetal bovine serum

GSK-3β:

glycogen synthase kinase-3β

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NMDA:

N-methyl-D-aspartate

PBS:

phosphate buffered saline

PI:

propidium iodide

TDZD-8:

4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione

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Correspondence to Zhuo Yang.

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Xu, J., Xu, P., Li, Z. et al. The role of glycogen synthase kinase-3β in glioma cell apoptosis induced by remifentanil. Cell Mol Biol Lett 18, 494–506 (2013). https://doi.org/10.2478/s11658-013-0102-3

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Key words

  • Remifentanil
  • Apoptosis
  • C6 cells
  • GSK-3β
  • TDZD-8
  • MTT
  • Hoechst 33342
  • Flow cytometry
  • NMDA receptor
  • Glioma