USP3 Promotes Glioma Progression by Stabilizing PLK1 through Deubiquitination

Glioma is an aggressive primary brain tumor with high morbidity and mortality, and it requires novel therapeutic targets. Ubiquitination and deubiquitination pathways regulate tumor progression. USP3, a deubiquitinating enzyme, and PLK1, a cell cycle kinase, have been linked to cancer cell proliferation. This study examined whether USP3 modulates PLK1 through deubiquitination to influence glioma progression. USP3 expression in glioma versus normal tissues was analyzed using TCGA datasets. Gain-of-function and loss-of-function models were generated by USP3 overexpression in U-251MG cells and USP3 knockdown in DK-MG cells. Proliferation was assessed by CCK-8, cell cycle distribution was analyzed by flow cytometry, and migration and invasion were evaluated using Transwell assays. Nude mouse xenografts were used to assess tumor growth, and PCNA immunohistochemistry was performed. Co-immunoprecipitation and Western blotting were used to validate the USP3-PLK1 interaction and to assess PLK1 ubiquitination. USP3 was upregulated in glioma tissues. USP3 overexpression enhanced U-251MG proliferation, promoted cell cycle progression, and increased invasion, while USP3 knockdown suppressed these phenotypes in DK-MG cells. Xenografts derived from USP3-overexpressing cells grew faster and showed higher PCNA levels, whereas USP3 knockdown reduced tumor growth and PCNA staining. Mechanistically, USP3 bound to PLK1, reduced K48-linked ubiquitination of PLK1, and stabilized PLK1 protein levels. USP3 knockdown increased PLK1 ubiquitination and reduced PLK1 abundance. Overall, USP3 promoted glioma malignancy by deubiquitinating and stabilizing PLK1, which enhanced proliferation and invasion. Targeting the USP3-PLK1 axis may represent a potential therapeutic strategy for glioma.