The effect of pseudouracil modifying enzyme 3 activating AKT pathway on malignant progression of glio-blastoma

ZHANG Chang; LIU Chunshan; LIAO Huaying; WU Yuchao; TIAN Yunhong

doi:10.3969/j.issn.1006-5725.2025.12.008

Abstract：Objective To investigate the regulatory effects and molecular mechanisms of PUS3 on GBM cell malignant behaviors(proliferation,apoptosis,invasion)in vitro,providing potential therapeutic targets for GBM.Methods The expression of PUS3 in GBM was analyzed using the GEPIA2 database.Kaplan-Meier survival analysis evaluated the survival difference between PUS3 high-and low-expression patients.qRT-PCR and Western blot were performed to detect PUS3 expression in normal glial cells(HEB)and GBM cell lines(U87,LN229,U251,T98G).PUS3-stably overexpressing GBM cell lines were constructed.Colony formation,CCK-8 assay,and flow cytometry were used to assess proliferation and apoptosis.Transwell assay evaluated cell invasion.Immunohis-tochemistry(IHC)detected PUS3 expression in GBM patient tissues.Western blot analyzed tumor-related pathway proteins after PUS3 overexpression.Results The expression level of PUS3 is elevated in GBM patient tissues,and the mRNA and protein expression levels in GBM cells are significantly higher than those in HEB cells(P<0.01).After overexpression of PUS3,the proliferation ability of GBM cells was enhanced(P<0.05),the apoptosis rate decreased(P<0.01),and the number of invasive cells increased(P<0.001).Mechanistically,overexpression of PUS3 significantly activates the AKT pathway,and the use of AKT inhibitors in PUS3 overexpressing cells can reverse the pro cancer effect.Conclusion PUS3 is highly expressed in glioblastoma(GBM)and promotes tumor cell proliferation,invasion,and apoptosis inhibition by activating the AKT pathway,suggesting its potential as a therapeutic target for GBM treatment.