Target proteomics-based analysis of the deep association between USP30 and prognosis of glioblastoma multiforme
Year: 2025
Type: article
Abstract: BACKGROUND AND PURPOSE: Glioblastoma Multiforme(GBM) is the most common primary malignant tumor of the central nervous system and the most prevalent primary malignant brain tumor in adults, accounting for over 50% of adult malignant primary brain tumors. Adult patients with GBM have a median age at onset ranging from 45.5 to 56 years, with no significant gender difference, and the 5-year survival rate is only 5%-10%. Despite the advancements in surgery and chemoradiotherapy, prognosis remains poor. There is an urgent need to identify novel therapeutic targets and prognostic biomarkers for GBM. Ubiquitin-specific protease 30 (USP30), a member of the deubiquitinating enzyme family, has been implicated in tumor progression. This study investigates the potential mechanisms of USP30 in GBM using proteomic analysis and functional validation. It further explores USP30's potential as a prognostic marker and therapeutic target in GBM, focusing on its downregulation and possible role in tumor suppression. METHODS: This study included paired tumor and adjacent tissues from six GBM patients. Differential protein expression was identified using LC-MS/MS (Liquid Chromatography-Tandem Mass Spectrometry, screening criteria: |log2FC| > 1 and P < 0.05). Functional enrichment analyses were performed using the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Reactome databases, and a protein-protein interaction network (confidence >0.7) was constructed via STRING to identify USP30 as a core molecule. In vitro functional experiments were conducted using U87 MG cells with USP30 overexpression (USP30 OE group). CCK-8, EdU, Transwell, colony formation, and flow cytometry assays were employed to validate USP30's regulatory effects on cell proliferation, migration, and cycle. RESULTS: LC-MS/MS identified 66 differentially expressed proteins in six pairs of GBM and adjacent tissues, with USP30 significantly downregulated in tumors (ratio = 0.54 ± 0.11, P = 0.0009). Functional enrichment analysis showed significant enrichment in "ubiquitin-like protein peptidase activity," "mitochondrial translation termination," and "ribosomal subunit" ( P < 0.001). KEGG pathways focused on "ribosome," "proteoglycans in cancer," and deubiquitination pathways. The protein interaction network revealed that USP30 formed core interaction modules with ubiquitination-related proteins (USP44 and JOSD2) and mitochondrial ribosomal proteins (MRPL33 and MRPL36; confidence >0.7).Functional experiments confirmed that USP30 overexpression significantly inhibited U87 cell proliferation (CCK-8 absorbance decreased by 44%, P = 6.0 × 10 -5 ), DNA synthesis (EdU-positive cells reduced by 38%, P = 0.0056), and colony formation (colony number decreased by 53%, P = 0.003764), while inducing G0/G1 cell cycle arrest (proportion increased by 65%, P < 0.001). Clinical validation showed that USP30 immunohistochemical scores in tumor tissues (1.20 ± 0.42) were significantly lower than those in adjacent tissues (2.53 ± 0.28, P < 0.0001). CONCLUSION: This study investigates the role of USP30 in GBM using proteomic analysis and in vitro functional assays. The study aims to explore USP30's potential as a prognostic marker and therapeutic target. Proteomic analysis revealed USP30's significant downregulation in GBM, which was further validated through in vitro experiments, where USP30 overexpression inhibited GBM cell proliferation and migration. These findings suggest that USP30 acts as a tumor suppressor in GBM, providing insights into its potential role in clinical applications. Further studies in larger cohorts are needed to confirm its prognostic relevance.
Source: International Journal of Surgery
Authors: Chao Wu, Yanfei Yang, Xiaoting Chang, Wei Shang
Language: English
Subfield: Genetics
Field: Medicine
Domain: Health Sciences
Open Access status: hybrid
Estimated APC paid: $3,450
OpenAlex ID: W4415913377