T52是从中国传统草药方块玉叶兰中提取出的一种甾体皂苷，据报道，它具有强烈的抗人咽喉癌细胞增殖能力。然而，T52是否具有抗骨肉瘤的特性及其潜在机制仍然未知。为了研究T52在骨肉瘤中的结果和潜在机制，我们使用CCK-8、集落形成、EdU染色、细胞周期/凋亡和细胞迁移/入侵实验来检查T52在骨肉瘤细胞中的生理作用。通过生物信息学预测评估T52与骨肉瘤相关的目标，并通过分子对接分析结合位点。进行西方印迹分析，检查与凋亡、细胞周期和STAT3信号通路激活相关的因子水平。T52在体外以剂量依赖的方式明显降低了骨肉瘤细胞的增殖、迁移和侵袭，并促进了G2/M阻滞和凋亡。在机理上，分子对接预测T52与STAT3 Src同源2（SH2）领域残基稳定结合。西方印迹显示，T52抑制了STAT3信号通路并减少了下游目标的表达，例如Bcl-2、Cyclin D1和c-Myc。此外，通过STAT3重新激活部分逆转T52的抗骨肉瘤性质，证实STAT3信号通路对调节T52的抗骨肉瘤性质至关重要。我们首次证明了T52在体外具有强大的抗骨肉瘤特性，这是通过抑制STAT3信号通路实现的。我们的发现为使用T52治疗骨肉瘤提供了药理支持，版权所有©2023 Elsevier GmbH发表。

T52 is a steroidal saponin extracted from the traditional Chinese herb Rohdea fargesii (Baill.), and it is reported to possess strong anti-proliferative capabilities in human pharyngeal carcinoma cell lines. However, whether T52 has anti-osteosarcoma properties, and its potential mechanism is remains unknown.To examine the outcome and underlying mechanism of T52 in osteosarcomas (OS).The physiological roles of T52 in OS cells were examined using CCK-8, colony formation (CF), EdU staining, cell cycle/apoptosis and cell migration/invasion assays. The relevant T52 targets against OS were assessed via bioinformatics prediction, and the binding sites were analyzed by molecular docking. Western blot analysis was carried out to examine the levels of factors associated with apoptosis, cell cycle, and STAT3 signaling pathway activation.T52 markedly diminished the proliferation, migration, and invasion of OS cells, and promoted G2/M arrest and apoptosis in a dose-dependent fashion (DDF) in vitro. Mechanistically, molecular docking predicted that T52 stably associated with STAT3 Src homology 2 (SH2) domain residues. Western blot revealed that T52 suppressed the STAT3 signaling pathway, as well as the expression of the downstream targets, such as, Bcl-2, Cyclin D1, and c-Myc. In addition, the anti-OS property of T52 were partially reversed by STAT3 reactivation, which confirmed that STAT3 signaling is critical for regulating the anti-OS property of T52.We firstly demonstrated that T52 possessed strong anti-osteosarcoma property in vitro, which was brought on by the inhibition of the STAT3 signaling pathway. Our findings provided pharmacological support for treating OS with T52.Copyright © 2023. Published by Elsevier GmbH.