恶性转化伴随着应激反应途径的过度激活。热休克蛋白（HSP）是应激诱导蛋白，在蛋白质折叠和加工中发挥作用，有助于应激肿瘤细胞的非癌基因成瘾。然而，HSP家族在骨肉瘤中的详细作用尚未得到研究。来自 GEO 和 TARGET 数据库的大量和单细胞转录组数据用于识别与骨肉瘤患者预后相关的 HSP。骨肉瘤中 HSPD1 的表达水平显着升高，与不良预后相关。通过体外和体内实验，我们系统地鉴定了HSPD1通过促进上皮间质转化（EMT）和激活AKT/mTOR信号传导来调节骨肉瘤的增殖、转移和凋亡。随后，使用免疫沉淀和质谱分析将 ATP5A1 确定为 HSPD1 的潜在靶标。从机制上讲，HSPD1可能与ATP5A1相互作用，减少K48相关的泛素化和ATP5A1的降解，最终激活AKT/mTOR通路，确保骨肉瘤进展和EMT过程。这些发现扩展了 HSPD1 发挥生物效应的潜在机制，并为其作为骨肉瘤的潜在治疗靶点提供了强有力的证据。© 作者。

Malignant transformation is concomitant with excessive activation of stress response pathways. Heat shock proteins (HSPs) are stress-inducible proteins that play a role in folding and processing proteins, contributing to the non-oncogene addiction of stressed tumor cells. However, the detailed role of the HSP family in osteosarcoma has not been investigated. Bulk and single-cell transcriptomic data from the GEO and TARGET databases were used to identify HSPs associated with prognosis in osteosarcoma patients. The expression level of HSPD1 was markedly increased in osteosarcoma, correlating with a negative prognosis. Through in vitro and in vivo experiments, we systematically identified HSPD1 as an important contributor to the regulation of proliferation, metastasis, and apoptosis in osteosarcoma by promoting the epithelial-mesenchymal transition (EMT) and activating AKT/mTOR signaling. Subsequently, ATP5A1 was determined as a potential target of HSPD1 using immunoprecipitation followed by mass spectrometry. Mechanistically, HSPD1 may interact with ATP5A1 to reduce the K48-linked ubiquitination and degradation of ATP5A1, which ultimately activates the AKT/mTOR pathway to ensure osteosarcoma progression and EMT process. These findings expand the potential mechanisms by which HSPD1 exerts biological effects and provide strong evidence for its inclusion as a potential therapeutic target in osteosarcoma.© The author(s).