Photodynamic therapy combined with temozolomide inhibits C6 glioma migration and invasion and promotes mitochondrial-associated apoptosis by inhibiting sodium-hydrogen exchanger isoform 1

Photodiagnosis Photodyn Ther. 2019 Jun:26:405-412. doi: 10.1016/j.pdpdt.2019.05.007. Epub 2019 May 11.

Abstract

Objective: As a targeted therapeutic technique for glioma inhibition, photodynamic therapy (PDT) has gradually become a focus of basic research related to glioma treatment. The capacity of PDT to kill glioma cells involves varieties of pathways. In glioma cells, activated sodium-hydrogen exchanger isoform 1 (NHE1) can inhibit the cytotoxic effect of temozolomide (TMZ), promote cell migration and invasion, and inhibit cell apoptosis by changing the acid-base equilibrium. The purpose of our study was to explore if PDT combined with TMZ can effectively inhibit glioma cells by influencing NHE1 in vitro.

Methods: We analyzed the expression levels of proteins such as NHE1, ezrin, vimentin, Bcl-2, and Bax by Western blot analysis, we assessed the migration and invasion of rat C6 glioma cells by Transwell assay, and we evaluated C6 cell apoptosis in vitro by flow cytometry.

Results: Western blot results indicated that NHE1, ezrin and vimentin were downregulated after cotreatment of C6 cells, and intracellular acidification was detected by a fluorometric intracellular pH assay. The migration and invasion capacities of C6 cells were significantly hindered after cotreatment, as shown by the Transwell assay. Experimental data also revealed a significant increase in cell apoptosis after cotreatment, as detected by flow cytometry; corresponding proapoptotic changes in Bcl-2, Bax and caspase-3 were also observed in vitro.

Conclusion: These results demonstrate that PDT combined with TMZ can inhibit C6 cell migration and invasion and promote mitochondrial-associated apoptosis by inhibiting NHE1. Therefore, this study provides supporting evidence for a potential method for the treatment of glioma.

Keywords: Glioma; Intracellular pH; Photodynamic therapy; Sodium-hydrogen exchanger isoform 1; Temozolomide.

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Caspase 3 / metabolism
  • Cell Movement / drug effects
  • Cytoskeletal Proteins / metabolism
  • Dose-Response Relationship, Drug
  • Glioma / drug therapy*
  • In Vitro Techniques
  • Mitochondria / drug effects
  • Photochemotherapy / methods*
  • Photosensitizing Agents / administration & dosage
  • Photosensitizing Agents / pharmacology*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Rats
  • Sodium-Hydrogen Exchanger 1 / metabolism
  • Temozolomide / administration & dosage
  • Temozolomide / pharmacology*
  • Tumor Cells, Cultured
  • Vimentin / metabolism
  • bcl-2-Associated X Protein / metabolism

Substances

  • Bax protein, rat
  • Bcl2 protein, rat
  • Cytoskeletal Proteins
  • Photosensitizing Agents
  • Proto-Oncogene Proteins c-bcl-2
  • SLC9A1 protein, human
  • Sodium-Hydrogen Exchanger 1
  • Vimentin
  • bcl-2-Associated X Protein
  • ezrin
  • Caspase 3
  • Temozolomide