iProXProteomicsHao ChenHomo Sapienshttp://www.iprox.org/page/project.html?id=IPX0006439000chenhao@sysucc.org.cnSun Yat-sen University Cancer CenteriProX<h4>Aims</h4>Esophageal squamous cell carcinoma (ESCC) is one of the aggressive and lethal malignancies with an extremely poor prognosis. It is necessary to explore the molecular mechanisms of ESCC invasion.<h4>Main methods</h4>We utilized high-throughput mass spectrometry to analyze the proteomes and phosphorylation profiles of two ESCC cell lines with differing invasion capacities (HK vs TE10). Differentially expressed proteins and phosphorites were identified, followed by comprehensive bioinformatics analyses encompassing function and pathway enrichment, protein-protein interaction (PPI) network analysis, hub gene identification, co-expression analysis, kinase-substrate prediction, and drug-target network analysis. CCK-8 assay, transwell examination, wound-healing assay, and western blot was used to validate the effects of fostamatinib on ESCC cells proliferation, invasion, migration, and LYN expression.<h4>Key findings</h4>The Q4 cluster of differentially phosphorylated proteins was primarily associated with functions and pathways relevant to tumor metastasis. Phosphorylated hub proteins including ARHGAP35, CTNNA1, and SHC1 were identified through the analysis of PPI network, and their respective regulated kinases were predicted. Among the predicted kinases, LYN was validated to be associated with lymph node metastasis (N0 vs. N1-3) and prognosis in ESCC patients at mRNA levels using TGGA data and protein levels in ESCC tissues (p < 0.05). Validation experiments confirmed the inhibitory effects of fostamatinib on ESCC cells proliferation, migration, invasion, and LYN expression.<h4>Significance</h4>Our multi-omics analysis offers deeper perspectives on ESCC invasiveness and unveils new phosphorylated hub proteins with their regulatory kinase. This study also suggests that fostamatinib may be a potential agent for treating ESCC.Integrated proteomics and phosphoproteomics analyses of esophageal cancer cells with different invasive abilities.Xu Nansong N, Lai Changchun C, He Qing-Mei QM, Cai Yubo Y, Yu Hui H, Zhong Wenhao W, Chen Shulin S, Wu Fang-Cai FC, Chen Hao HfalseIntegrated proteomics and phosphoproteomics analyses of esophageal cancer cell with different invasive abilitiestwo of the ten esophageal cancer cells with the strongest and weakest invasiveness were selected for proteomics and phosphorylation proteomics analysis to screen differential phosphorylation sites related to cell invasionMon May 22 00:00:00 GMT+01:00 2023PXD042408960637734435