iProXProteomicsHao GuHomo Sapienshttp://www.iprox.org/page/project.html?id=IPX0012686000guhao@ahmu.edu.cnAnhui Medical UniversityiProX<h4>Background</h4>Portal vein tumor thrombus is a lethal complication of HCC for which effective biomarkers are urgently needed.<h4>Methods</h4>Through proteomic profiling, functional and mechanistic assays, and machine learning-based evaluation of serum NSUN6 in 472 HCC patients with external validation (n = 98), we characterized an epitranscriptomic regulator of HCC metastasis.<h4>Results</h4>NSUN6 was downregulated in PVTT and suppressed metastasis by catalyzing m⁵C modification of SAV1 mRNA, stabilizing its transcript and activating Hippo signaling. Furthermore, NSUN6 was released from HCC cells via extracellular vesicle-associated and non-canonical pathways. Circulating NSUN6 levels were inversely associated with metastasis in both discovery and validation cohorts, and lower levels predicted shorter metastasis-free survival.<h4>Conclusion</h4>Our findings establish the NSUN6-SAV1-YAP axis as a key epitranscriptomic tumor-suppressive mechanism and support circulating NSUN6 as a potential biomarker for early risk stratification in HCC.Circulating NSUN6 serves as a prognostic biomarker for early metastasis in HCC via hippo pathway modulation.Li Xutong X, Xu Honghai H, Chang Yu Y, Zhang Yani Y, Xu Hui H, Yin Xin X, Zhang Wanxiang W, Luo Yi Y, Lu Wanting W, Fan Wenhu W, Ruan Wanwan W, Shen Jiapei J, Li Zhenxing Z, Du Shucheng S, Bentum-Ennin Lutterodt L, Ni Jing J, Gao Wei W, Li Jiabin J, Hu Wanglai W, Yang Wanshui W, Gao Yufeng Y, Gu Hao HfalseO-GlcNAc Glycosylation 4D-Label free Quantitative Proteomics Bioinformatics Analysis ReportO-linked β-N-acetylglucosamine (O-GlcNAc) modification is a dynamic post-translational modification that involves the reversible attachment of N-acetylglucosamine to serine or threonine residues of nuclear, cytoplasmic, and mitochondrial proteins. This modification is critical for regulating cellular processes such as signal transduction, transcription, translation, protein degradation, and cellular localization. Aberrant O-GlcNAcylation has been implicated in numerous pathological conditions, including diabetes, cardiovascular diseases, neurodegenerative disorders, and cancers, highlighting the need for precise methods to study its dynamics across biological systems.Wed Nov 26 00:00:00 GMT 2025PXD066348960642464346