Sp1 Upregulation Bolsters the Radioresistance of Glioblastoma Cells by Promoting Double Strand Breaks Repair
Abstract
:1. Introduction
2. Results
2.1. IR Upregulated Sp1 Expression in GBM Cells
2.2. IR Upregulated Sp1 Expression in GBM Cells In Vivo
2.3. Sp1 Knockdown Radiosensitized GBM Cells In Vitro
2.4. Sp1 Knockdown Radiosensitized GBM Cells In Vivo
2.5. Sp1 Activated the DNA-PKcs Promoter and Enhanced DNA-PKcs Expression and Activity
2.6. p-Sp1 Was Located in the DSB Sites
2.7. Sp1 Silencing Delayed IR-Induced DSB Repair
2.8. Sp1 Silencing Aggravated IR-Induced Apoptosis In Vivo and In Vitro
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Culture
4.2. Irradiation
4.3. RNA Extraction and RT-PCR
4.4. siRNA Transfection and Establishment of Stable Sp1-Knockdown Cell Lines
4.5. Western Blotting
4.6. Immunofluorescence
4.7. Dual Luciferase Assay
4.8. Clonogenic Assay
4.9. Apoptosis Analysis
4.10. Xenograft Model
4.11. Bioinformatics Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
GBM | Glioblastoma |
DSB | Double strand breaks |
Sp1 | Specificity protein 1 |
IR | Ionizing radiation |
DNA-PKcs | DNA-dependent protein kinase catalytic subunit |
RT-PCR | Reverse transcription-polymerase chain reaction |
FBS | Fetal bovine serum |
ATM | Ataxia-telangiectasia mutated |
DDB | Damage-specific DNA-binding protein |
NFDB1 | Nuclear factor with BRCT domain 1 |
MDC1 | Mediator of DNA damage checkpoint protein 1 |
XRCC1 | X-ray repair cross-complementing gene 1 |
NHEJ | Non-homologous end joining |
HR | Homologous recombination |
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Total Apoptotic Cells (%) | ||
---|---|---|
U87 | M059K | |
NC | 6.67 ± 1.26 | 5.76 ± 0.56 |
si-SP1 | 6.20 ± 0.87 | 6.26 ± 0.48 |
NC + IR | 19.72 ± 2.79 | 15.03 ± 2.07 |
si-Sp1 + IR | 49.57 ± 8.71 ***### | 48.53 ± 9.93 ***### |
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Liu, X.; Sun, C.; Wang, Q.; Li, P.; Zhao, T.; Li, Q. Sp1 Upregulation Bolsters the Radioresistance of Glioblastoma Cells by Promoting Double Strand Breaks Repair. Int. J. Mol. Sci. 2023, 24, 10658. https://doi.org/10.3390/ijms241310658
Liu X, Sun C, Wang Q, Li P, Zhao T, Li Q. Sp1 Upregulation Bolsters the Radioresistance of Glioblastoma Cells by Promoting Double Strand Breaks Repair. International Journal of Molecular Sciences. 2023; 24(13):10658. https://doi.org/10.3390/ijms241310658
Chicago/Turabian StyleLiu, Xiongxiong, Chao Sun, Qiqi Wang, Ping Li, Ting Zhao, and Qiang Li. 2023. "Sp1 Upregulation Bolsters the Radioresistance of Glioblastoma Cells by Promoting Double Strand Breaks Repair" International Journal of Molecular Sciences 24, no. 13: 10658. https://doi.org/10.3390/ijms241310658