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Published on: June 12, 2019
Tetraarsenic oxide-induced inhibition of malignant glioma cell invasion in vitro via a decrease in matrix
Ho-Shin Gwak1, Myung-Jin Park, In-Chul Park
1Neuro-Oncology Clinic, National Cancer Center, Goyang;
Object:
Local invasiveness of malignant glioma is a major reason for the failure of current treatments including surgery and radiation therapy. Tetraarsenic oxide (As4O6 [TAO]) is a trivalent arsenic compound that has potential anticancer and antiangiogenic effects in selected cancer cell lines at a lower concentration than arsenic trioxide (As2O3 [ATO]), which has been more widely tested in vitro and in vivo. The authors tried to determine the cytotoxic concentration of TAO in malignant glioma cell lines and whether TAO would show anti-invasive effects under conditions independent of cell death or apoptosis.
Methods:
The human phosphatase and tensin homolog (PTEN)-deficient malignant glioma cell lines U87MG, U251MG, and U373MG together with PTEN-functional LN428 were cultured with a range of micromolar concentrations of TAO. The invasiveness of the glioma cell lines was analyzed. The effect of TAO on matrix metalloproteinase (MMP) secretion and membrane type 1 (MT1)-MMP expression was measured using gelatin zymography and Western blot, respectively. Akt, or protein kinase B, activity, which is a downstream effector of PTEN, was assessed with a kinase assay using glycogen synthesis kinase-3β (GSK-3β) as a substrate and Western blotting of phosphorylated Akt.
Results:
Tetraarsenic oxide inhibited 50% of glioma cell proliferation at 6.3-12.2 μM. Subsequent experiments were performed under the same TAO concentrations and exposure times, avoiding the direct tumoricidal effect of TAO, which was confirmed with apoptosis markers. An invasion assay revealed a dose-dependent decrease in invasiveness under the influence of TAO. Both the gelatinolytic activity of MMP-2 and MT1-MMP expression decreased in a dose-dependent manner in all cell lines, which was in accordance with the invasion assay results. The TAO decreased kinase activity of Akt on GSK-3β assay and inhibited Akt phosphorylation in a dose-dependent manner in all cell lines regardless of their PTEN status.
Conclusions:
These results showed that TAO effectively inhibits proliferation of glioblastoma cell lines and also exerts an anti-invasive effect via decreased MMP-2 secretion, decreased MT1-MMP expression, and the inhibition of Akt phosphorylation under conditions devoid of cytotoxicity. Further investigations using an in vivo model are needed to evaluate the potential role of TAO as an anti-invasive agent.
Insights
Tetraarsenic oxide (TAO) inhibits malignant glioma cell proliferation and invasiveness by reducing MMP-2, MT1-MMP, and Akt phosphorylation. This study explores TAO
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Malignant glioma's local invasiveness hinders treatment efficacy.
- Tetraarsenic oxide (TAO) shows potential anticancer effects at lower concentrations than arsenic trioxide (ATO).
- TAO's anti-invasive properties and cytotoxic concentration in glioma require elucidation.
Purpose of the Study:
- Determine the cytotoxic concentration of TAO in malignant glioma cell lines.
- Investigate TAO's anti-invasive effects independent of cell death.
- Assess TAO's impact on matrix metalloproteinase (MMP) and Akt signaling pathways.
Main Methods:
- Cultured PTEN-deficient (U87MG, U251MG, U373MG) and PTEN-functional (LN428) glioma cells with varying TAO concentrations.
- Analyzed cell invasiveness, MMP secretion (gelatin zymography), MT1-MMP expression (Western blot), and Akt activity (kinase assay, Western blot).
Main Results:
- TAO inhibited 50% of glioma cell proliferation at 6.3-12.2 μM.
- TAO significantly reduced cell invasiveness, MMP-2 activity, and MT1-MMP expression in a dose-dependent manner.
- TAO inhibited Akt phosphorylation and kinase activity irrespective of PTEN status.
Conclusions:
- TAO effectively inhibits glioblastoma cell proliferation and invasiveness.
- TAO's anti-invasive mechanism involves decreased MMP-2 secretion, MT1-MMP expression, and Akt phosphorylation, independent of cytotoxicity.
- Further in vivo studies are warranted to explore TAO's therapeutic potential as an anti-invasive agent.
