Knocking down RFWD3 suppresses the growth and migration of glioblastoma cells by blocking G2/M cycle progression.

Glioblastoma (GBM) is the most common and lethal primary malignant tumor of the adult central nervous system, accounting for approximately half of all gliomas. Although RING finger and WD repeat domain 3 (RFWD3) has been implicated in the regulation of various cancers, its role and underlying mechanisms in GBM progression remain unclear. In this study, analysis of the GEPIA online database confirmed that RFWD3 expression is significantly elevated in GBM tumor tissues. We found increased RFWD3 protein levels in U87 and T98 GBM cell lines and that suppression of RFWD3 markedly reduced cell viability and increased apoptosis in the GBM lines. Moreover, RFWD3 knockdown significantly inhibited their migration and invasion. Flow cytometry analysis revealed that RFWD3 inhibition induced G2/M cell cycle arrest (U87: G1 phase from 69.03% to 50.48%, G2/M phase from 16.54% to 33.01%; T98: G1 phase from 67.31% to 48.32%, G2/M phase from 17.10% to 33.73%). Furthermore, suppression of RFWD3 downregulated the Wnt/β-catenin signaling pathway. These findings indicate that RFWD3 promotes GBM cell growth and migration by facilitating G2/M phase progression through the Wnt/β-catenin pathway. Targeting RFWD3 may represent a promising strategy for inhibiting GBM progression.