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Inhibitory effect of pisosterol on human glioblastoma cell lines with C-MYC amplification
Edmundo L R Pereira1, Patrícia D L Lima, André S Khayat
1Human Cytogenetics Laboratory, Institute of Biological Sciences, Federal University of Pará, Av. Augusto Corrêa, 01, CEP 66075-110 - Belém/PA, Brazil.
Abstract:
Despite the remarkable progress in the characterization of the molecular pathogenesis of glioblastoma multiforme (GBM), these tumors remain incurable and, in most cases, refractory to aggressive cytotoxic treatments. We conducted a morphological and cytogenetic study in two GBM cell lines (U343 and AHOL1), before and after treatment with pisosterol (at 0.5, 1.0 and 1.8 µg ml⁻¹), a triterpene isolated from the fungus Pisolithus tinctorius. No significant alteration was observed in the morphology and frequency of chromosomal abnormalities in the cell lines analyzed after treatment with pisosterol. Using fluorescence in situ hybridization analysis with a locus-specific probe for C-MYC showed that 72% of U343 and 65% of AHOL1 cells contained more than two alleles of C-MYC before treatment. After treatment, no effects were detected at lower concentrations of pisosterol (0.5 and 1.0 µg ml⁻¹). However, at 1.8 µg ml⁻¹ of pisosterol, only 33% of U343 cells and 15% of AHOL1 cells presented more than two fluorescent signals, suggesting that pisosterol blocks the cells with gene amplification. Cells that do not show a high degree of C-MYC gene amplification have a less aggressive and invasive behavior and are easy targets for chemotherapy. Therefore, further studies are needed to examine the use of pisosterol in combination with conventional anti-cancer therapy.
Insights
Pisosterol, a compound from Pisolithus tinctorius, reduced C-MYC gene amplification in glioblastoma multiforme (GBM) cells. This suggests potential for combining pisosterol with chemotherapy for treating this incurable brain cancer.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Glioblastoma multiforme (GBM) remains largely incurable despite advances in understanding its molecular pathogenesis.
- Current cytotoxic treatments are often ineffective against GBM tumors.
Purpose of the Study:
- To investigate the effects of pisosterol, a triterpene from Pisolithus tinctorius, on GBM cell lines.
- To assess pisosterol's impact on C-MYC gene amplification in U343 and AHOL1 GBM cells.
Main Methods:
- Morphological and cytogenetic analysis of U343 and AHOL1 GBM cell lines.
- Treatment with varying concentrations of pisosterol (0.5, 1.0, 1.8 µg ml⁻¹).
- Fluorescence in situ hybridization (FISH) to analyze C-MYC gene copy number.
Main Results:
- Pisosterol did not significantly alter cell morphology or the frequency of chromosomal abnormalities.
- At 1.8 µg ml⁻¹, pisosterol reduced C-MYC gene amplification in both U343 (72% to 33%) and AHOL1 (65% to 15%) cells.
- Lower pisosterol concentrations (0.5 and 1.0 µg ml⁻¹) showed no effect on C-MYC amplification.
Conclusions:
- Pisosterol, at a concentration of 1.8 µg ml⁻¹, effectively blocks C-MYC gene amplification in GBM cells.
- Reduced C-MYC amplification is associated with less aggressive GBM behavior and increased chemosensitivity.
- Pisosterol warrants further investigation for combination therapy with conventional anti-cancer treatments for GBM.
