LARS promotes osteosarcoma proliferation through leucine-dependent PRIM2 translation and DNA replication activation

Osteosarcoma (OS) is an aggressive bone malignancy in adolescents, with poor prognosis and limited survival improvement over decades, necessitating new therapeutic targets. Prior research identified Leucyl-tRNA synthetase (LARS) as critical for OS proliferation, prompting this investigation into its underlying mechanisms. Utilizing clinical OS samples, we assessed LARS and primase p58 subunit 2 (PRIM2) expression via immunohistochemistry. In vitro studies employed OS cell lines for LARS/PRIM2 overexpression or knockdown, followed by functional assays: MTT, colony formation, EdU staining, Transwell migration/invasion, and flow cytometry for cell cycle/ROS/Ca2⁺ analysis. Xenograft models were used to evaluate tumor progression in vivo. Multi-omics analyses included transcriptome sequencing, proteomic profiling, and telomeric repeat amplification protocol-PCR to assess translational regulation. Stable isotope labeling by amino acids in cell culture (SILAC) determined leucine-dependent PRIM2 synthesis. Mechanisms were further probed using inhibitors and rescue experiments. LARS expression is significantly elevated in OS, and its overexpression enhances proliferation but inhibits invasion and migration in vitro and in vivo. Conversely, LARS silencing in OS cells results in cell cycle arrest. Mechanistically, the upregulation of LARS in OS is associated with increased glycolysis and DNA replication and a reduction in endoplasmic reticulum stress, while elevating the oncogene PRIM2 through leucine-dependent translational control. Our findings highlight the crucial oncogenic role of the LARS/PRIM2 axis in promoting the pathogenesis of OS, primarily through the alleviation of endoplasmic reticulum stress and the activation of translation processes.