PXDN regulated by WTAP/YTHDF1-mediated m(6)A modification activates PI3K/AKT signaling pathway through extracellular matrix remodeling to promote progression in nasopharyngeal carcinoma

PXDN受WTAP/YTHDF1介导的m(6)A修饰调控,通过细胞外基质重塑激活PI3K/AKT信号通路,促进鼻咽癌的进展。

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Abstract

BACKGROUND: Recurrence and metastasis remain the primary causes of treatment failure in nasopharyngeal carcinoma (NPC). This study aims to explore the functional role and regulatory mechanisms of peroxidasin (PXDN) in NPC progression. METHODS: Weighted gene co-expression network analysis was performed to screen the module most relevant to the malignant progression of NPC from our internal cohort (Fujian cohort 1, N = 192), from which the PXDN was identified as the key molecule. Clinical significance of PXDN was assessed in the GEO database and NPC tissue microarrays from Fujian cohort 2 (N = 103). Functional experiments were used to determine the biological role of PXDN in NPC. Methylated RNA immunoprecipitation sequencing was used to identify PXDN N6-methyladenosine (m(6)A) modification site, and verified by a dual-luciferase reporter gene assay. RNA immunoprecipitation, RNA stability assay, and RNA pull-down assay were used to verify the relationship between PXDN and YTHDF1. The CRISPR/Cas9 system was used to disrupt the m⁶A motif in the PXDN gene to further validate its role. Next, transcriptome, proteomic analysis, and immunoprecipitation assay were conducted to assess downstream targets. RESULTS: PXDN was highly expressed in NPC and associated with poor prognosis. Suppression of PXDN drastically reduced proliferation, migration, invasion, and resistance to chemoradiation in vitro, concomitant with attenuated epithelial-mesenchymal transition. Knockdown of PXDN remarkably suppressed NPC tumorigenicity and liver metastasis in vivo. Mechanistically, PXDN promotes aggressiveness by extracellular matrix remodelling to activate the ITGB1-PI3K-AKT pathway. The aberrant expression of PXDN is governed by an m(6)A-based regulatory axis, wherein YTHDF1 recognizes WTAP-mediated PXDN m(6)A methylation to enhance its RNA stability and translation. Targeted specific demethylation of PXDN m(6)A by CRISPR/Cas9 system significantly decreased the expression of PXDN. CONCLUSIONS: We reveal the crucial role of PXDN in driving NPC malignancy and the regulatory role of m(6)A methylation modification. Targeting PXDN expression or activity could be used to effectively control NPC progression. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-025-03609-y.

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