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Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance
Gaia Pucci1,2, Luigi Minafra1,2, Valentina Bravatà1
1Institute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, Italy.
Abstract:
The hypoxic pattern of glioblastoma (GBM) is known to be a primary cause of radioresistance. Our study explored the possibility of using gene knockdown of key factors involved in the molecular response to hypoxia, to overcome GBM radioresistance. We used the U87 cell line subjected to chemical hypoxia generated by CoCl2 and exposed to 2 Gy of X-rays, as single or combined treatments, and evaluated gene expression changes of biomarkers involved in the Warburg effect, cell cycle control, and survival to identify the best molecular targets to be knocked-down, among those directly activated by the HIF-1α transcription factor. By this approach, glut-3 and pdk-1 genes were chosen, and the effects of their morpholino-induced gene silencing were evaluated by exploring the proliferative rates and the molecular modifications of the above-mentioned biomarkers. We found that, after combined treatments, glut-3 gene knockdown induced a greater decrease in cell proliferation, compared to pdk-1 gene knockdown and strong upregulation of glut-1 and ldha, as a sign of cell response to restore the anaerobic glycolysis pathway. Overall, glut-3 gene knockdown offered a better chance of controlling the anaerobic use of pyruvate and a better proliferation rate reduction, suggesting it is a suitable silencing target to overcome radioresistance.
Insights
Targeting glucose transporter 3 (GLUT-3) gene knockdown in glioblastoma (GBM) can reduce radioresistance. Silencing GLUT-3 effectively controls pyruvate use and decreases proliferation, offering a promising strategy against GBM.
Area of Science:
- Oncology
- Molecular Biology
- Radiotherapy Research
Background:
- Hypoxia in glioblastoma (GBM) significantly contributes to radioresistance.
- Targeting molecular responses to hypoxia is a potential strategy to enhance GBM treatment efficacy.
Purpose of the Study:
- To investigate gene knockdown of hypoxia-inducible factors to overcome GBM radioresistance.
- To identify optimal molecular targets for gene silencing among HIF-1α activated factors.
Main Methods:
- Utilized U87 glioblastoma cells under chemical hypoxia (CoCl2) and X-ray irradiation (2 Gy).
- Evaluated gene expression of biomarkers related to the Warburg effect, cell cycle, and survival.
- Employed morpholino-induced gene silencing for selected genes, including glut-3 and pdk-1.
- Assessed cell proliferation rates and molecular modifications post-gene silencing.
Main Results:
- Glut-3 gene knockdown demonstrated a greater reduction in cell proliferation compared to pdk-1 knockdown.
- Observed significant upregulation of glut-1 and ldha following combined treatments, indicating a cellular response to restore anaerobic glycolysis.
- Glut-3 silencing effectively controlled pyruvate metabolism and reduced proliferation rates.
Conclusions:
- Glut-3 gene knockdown is a promising strategy for controlling anaerobic pyruvate metabolism in glioblastoma.
- Targeting glut-3 offers a viable approach to reduce glioblastoma proliferation and overcome radioresistance.
