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Phospho-Proteomics Analysis of Early Response to X-Ray Irradiation Reveals Molecular Mechanism Potentially Related to
Ousseynou Ben Diouf1, Antoine Gilbert2, Benoit Bernay3
1Mixed Research Exploration and Diagnosis (UMRED), UFR-Healthy, Iba Der THIAM University of Thies, Thies BP A967, Senegal.
Abstract:
Glioblastoma (GBM) is a devastating malignant brain tumor with a poor prognosis. GBM is associated with radioresistance. Post-translational modifications (PTMs) such as protein phosphorylation can play an important role in the cellular response to radiation. To better understand the early cellular activities after radiation in GBM, we carried out a phospho-proteomic study on the U251 cell line 3 h after X-ray irradiation (6Gy) and on non-irradiated cells. Our study showed a strong modification of proteoform phosphorylation in response to radiation. We found 453 differentially expressed phosphopeptides (DEPs), with 211 being upregulated and 242 being downregulated. A GO enrichment analysis of DEPs showed a strong enrichment of the signaling pathways involved in DNA damage response after irradiation and categorized them into biological processes (BPs), cellular components (CCs) and molecular functions (MFs). Certain accessions such as BRCA1, MDC1, H2AX, MDC1, TP53BP1 were dynamically altered in our fraction and are highly associated with the signaling pathways enriched after radiation.
Insights
This study reveals how protein phosphorylation changes in glioblastoma cells soon after radiation exposure. These findings offer insights into radioresistance mechanisms in brain tumors.
Area of Science:
- Oncology
- Molecular Biology
- Proteomics
Background:
- Glioblastoma (GBM) is an aggressive brain tumor with limited treatment options.
- GBM exhibits significant radioresistance, complicating radiotherapy outcomes.
- Protein phosphorylation, a key post-translational modification, influences cellular responses to radiation.
Purpose of the Study:
- To investigate early changes in protein phosphorylation in GBM cells following X-ray irradiation.
- To identify specific signaling pathways affected by radiation in GBM.
Main Methods:
- Phosphoproteomic analysis was performed on the U251 glioblastoma cell line.
- Cells were analyzed 3 hours post-irradiation (6 Gy) and compared to non-irradiated controls.
- Bioinformatic analysis, including Gene Ontology (GO) enrichment, was used to interpret phosphoproteomic data.
Main Results:
- A significant alteration in proteoform phosphorylation was observed in response to radiation.
- 453 differentially expressed phosphopeptides (DEPs) were identified, with 211 upregulated and 242 downregulated.
- GO enrichment analysis highlighted pathways involved in DNA damage response, including key proteins like BRCA1, H2AX, and TP53BP1.
Conclusions:
- Radiation exposure rapidly induces widespread changes in protein phosphorylation in GBM cells.
- These phosphorylation dynamics are strongly linked to DNA damage response pathways.
- Understanding these early events may provide targets for overcoming radioresistance in glioblastoma.
