骨肉瘤进展中的免疫微环境：PLEK 作为预后生物标志物和治疗靶点

We conducted differential expression and survival analyses using OS transcriptomic datasets and TCGA/GTEx data. Protein-protein interaction networks, GO/KEGG enrichment, and CytoHubba algorithms identified core hub genes. Tumor-infiltrating immune cells were profiled via TIMER. Single-cell RNA-seq (GSE162454) was used for immune and metabolic landscape mapping. PLEK was further validated by qRT-PCR and Western blot in OS samples, and its function assessed via siRNA knockdown in macrophages within TME co-cultured with OS cells. Cell proliferation, migration, and invasion assays evaluated phenotypic effects in OS cells.

Nine hub genes were identified, with PLEK significantly upregulated in OS tissues. High PLEK expression correlated with improved survival and increased infiltration of macrophages, dendritic cells, and CD4 + T cells. Single-cell analysis showed PLEK enrichment in macrophage-dominated clusters with active glycolytic and oxidative phosphorylation pathways. Downregulation of PLEK in macrophages enhanced OS cell proliferation, migration and invasion. These findings suggest PLEK is linked to a pro-immune, metabolically active microenvironment and may act as a tumor suppressor. DISCUSSION: Our study identifies PLEK as a prognostic biomarker and functional regulator in OS. It promotes an immune-infiltrated, metabolically active tumor microenvironment and is associated with attenuated malignant phenotypes in vitro . These findings highlight PLEK as a promising target for immunometabolic modulation in OS.