AMPA receptor antagonist perampanel affects glioblastoma cell growth and glutamate release in vitro

PLoS One. 2019 Feb 4;14(2):e0211644. doi: 10.1371/journal.pone.0211644. eCollection 2019.

Abstract

Epileptic seizures are frequent in patients with glioblastoma, and anticonvulsive treatment is often necessary. While clinical guidelines recommend all approved anticonvulsants, so far it is still unclear which of the available drugs is the best therapeutic option for treating glioma-associated seizures, also in view of possible anti-tumorigenic effects. In our study, we employed four patient-derived low-passage cell lines of glioblastoma and three cell lines of brain metastases, and challenged these cultures with four anticonvulsants with different mechanisms of action: levetiracetam, valproic acid, carbamazepine and perampanel. Cell proliferation was determined by bromodeoxyuridine incorporation. To further analyze the effects of perampanel, apoptosis induction was measured by caspase 3/7 activation. Glutamate release was quantified and glucose uptake was determined using 18F-fluorodeoxyglucose. Real-time polymerase chain reaction was employed to assess the expression of genes associated with glutamate release and uptake in brain tumor cells. Of the four anticonvulsants, only perampanel showed systematic inhibitory effects on cell proliferation, whereas all other anticonvulsants failed to inhibit glioma and metastasis cell growth in vitro. Metastasis cells were much more resistant to perampanel than glioblastoma cell lines. Glucose uptake was attenuated in all glioblastoma cells after perampanel exposure, whereas cell death via apoptosis was not induced. Extracellular glutamate levels were found to be significantly higher in glioblastoma cell lines as compared to metastasis cell lines, but could be reduced by perampanel exposure. Incubation with perampanel up-regulated glutamine synthetase expression in glioblastoma cells, whereas treatment with valproic acid and levetiracetam downregulated excitatory amino acid transporter-2 expression. Overall, our data suggest that perampanel acts as an anticonvulsive drug and additionally mediated anti-tumorigenic effects.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anticonvulsants / pharmacology
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Brain / drug effects
  • Brain / metabolism
  • Brain Neoplasms / drug therapy*
  • Brain Neoplasms / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / drug effects*
  • Down-Regulation / drug effects
  • Epilepsy / drug therapy
  • Epilepsy / metabolism
  • Glioblastoma / drug therapy*
  • Glioblastoma / metabolism
  • Glutamic Acid / metabolism*
  • Humans
  • Pyridones / pharmacology*
  • Receptors, AMPA / antagonists & inhibitors*
  • Seizures / drug therapy
  • Seizures / metabolism
  • Up-Regulation / drug effects
  • Valproic Acid / pharmacology

Substances

  • Anticonvulsants
  • Antineoplastic Agents
  • Pyridones
  • Receptors, AMPA
  • Glutamic Acid
  • Valproic Acid
  • perampanel

Grant support

This work was supported by a grant from the University Medicine Rostock FORUN Program to F.L. (Project no.: 889010) and a grant by the Damp Stiftung (Project no.:2016-20) to F.L. and T.K. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.