Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance
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 hypox...
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MDPI AG
2024-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/4/2079 |
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| author | Gaia Pucci Luigi Minafra Valentina Bravatà Marco Calvaruso Giuseppina Turturici Francesco P. Cammarata Gaetano Savoca Boris Abbate Giorgio Russo Vincenzo Cavalieri Giusi I. Forte |
| author_facet | Gaia Pucci Luigi Minafra Valentina Bravatà Marco Calvaruso Giuseppina Turturici Francesco P. Cammarata Gaetano Savoca Boris Abbate Giorgio Russo Vincenzo Cavalieri Giusi I. Forte |
| author_sort | Gaia Pucci |
| collection | DOAJ |
| description | 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, <i>glut-3</i> and <i>pdk-1</i> 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, <i>glut-3</i> gene knockdown induced a greater decrease in cell proliferation, compared to <i>pdk-1</i> gene knockdown and strong upregulation of <i>glut-1</i> and <i>ldha</i>, as a sign of cell response to restore the anaerobic glycolysis pathway. Overall, <i>glut-3</i> 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. |
| first_indexed | 2024-03-07T22:29:52Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-07T22:29:52Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-94a2dd7e41c94bb59b13744556b843e82024-02-23T15:19:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01254207910.3390/ijms25042079Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma RadioresistanceGaia Pucci0Luigi Minafra1Valentina Bravatà2Marco Calvaruso3Giuseppina Turturici4Francesco P. Cammarata5Gaetano Savoca6Boris Abbate7Giorgio Russo8Vincenzo Cavalieri9Giusi I. Forte10Institute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, ItalyInstitute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, ItalyInstitute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, ItalyInstitute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, ItalyDepartment of Biological, Chemical and Pharmaceutical Sciences and Technologies (STeBiCeF), University of Palermo, Viale delle Scienze Bld.17, 90128 Palermo, ItalyInstitute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, ItalyRadiation Oncology, ARNAS-Civico Hospital, 90100 Palermo, ItalyRadiation Oncology, ARNAS-Civico Hospital, 90100 Palermo, ItalyInstitute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, ItalyDepartment of Biological, Chemical and Pharmaceutical Sciences and Technologies (STeBiCeF), University of Palermo, Viale delle Scienze Bld.17, 90128 Palermo, ItalyInstitute of Molecular Bioimaging and Physiology (IBFM)-National Research Council (CNR), Cefalù Secondary Site, C/da Pietrapollastra-Pisciotto, 90015 Cefalù, ItalyThe 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, <i>glut-3</i> and <i>pdk-1</i> 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, <i>glut-3</i> gene knockdown induced a greater decrease in cell proliferation, compared to <i>pdk-1</i> gene knockdown and strong upregulation of <i>glut-1</i> and <i>ldha</i>, as a sign of cell response to restore the anaerobic glycolysis pathway. Overall, <i>glut-3</i> 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.https://www.mdpi.com/1422-0067/25/4/2079glioblastomaradioresistancechemical hypoxiagene knockdown |
| spellingShingle | Gaia Pucci Luigi Minafra Valentina Bravatà Marco Calvaruso Giuseppina Turturici Francesco P. Cammarata Gaetano Savoca Boris Abbate Giorgio Russo Vincenzo Cavalieri Giusi I. Forte Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance International Journal of Molecular Sciences glioblastoma radioresistance chemical hypoxia gene knockdown |
| title | Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance |
| title_full | Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance |
| title_fullStr | Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance |
| title_full_unstemmed | Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance |
| title_short | Glut-3 Gene Knockdown as a Potential Strategy to Overcome Glioblastoma Radioresistance |
| title_sort | glut 3 gene knockdown as a potential strategy to overcome glioblastoma radioresistance |
| topic | glioblastoma radioresistance chemical hypoxia gene knockdown |
| url | https://www.mdpi.com/1422-0067/25/4/2079 |
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