3-aminopyridine-2-carboxogenic thiosemicarbazone (3-AP) 具有广谱抗肿瘤活性。然而，其在骨肉瘤（OS）中的作用仍不清楚。因此，本研究利用三种人OS细胞系（MG-63、U2-OS和143B）以及移植143B细胞产生的裸鼠模型，探讨了3-AP对体外和体内OS的影响。用 DMSO（对照）或梯度浓度的 3-AP 处理细胞和小鼠。然后，进行各种测定（例如细胞计数试剂盒8、流式细胞术、免疫组织化学和蛋白质印迹）以评估细胞活力和凋亡水平，以及γH2A.X（DNA损伤相关性）、核糖核苷酸还原酶催化亚基M1和M2（分别为 RRM1 和 RRM2）蛋白质水平（铁依赖性相关性）。 3-AP 可以时间和剂量依赖性地抑制所有三种 OS 细胞系的生长并诱导细胞凋亡，柠檬酸铁铵 (FAC) 可以阻断这些作用。此外，3-AP降低了RRM2和总核糖核苷酸还原酶（RRM1​​加RRM2）蛋白的表达，但显着增加了γH2A.X的表达；治疗不影响 RRM1 表达。 FAC 治疗再次减弱了这些影响。在体内，与对照小鼠相比，3-AP处理的小鼠肿瘤切片中凋亡细胞的数量增加。 3-AP 处理还降低了 Ki-67 和 p21 的表达，表明抑制了 OS 生长。此外，RRM1、RRM2 和转铁蛋白受体蛋白 1（即 Tfr1）的表达表明 3-AP 通过铁依赖性途径抑制 OS 生长。总之，3-AP 通过降低铁依赖性途径的活性在 OS 中表现出抗癌活性，这可能是 OS 的一种有前途的治疗策略。© 2023。作者，获得 Springer Science Business Media, LLC 的独家许可，施普林格自然的一部分。

3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP) has broad-spectrum antitumor activity. However, its role in osteosarcoma (OS) remains unclear. Therefore, this study explored the effects of 3-AP on OS in vitro and in vivo using three human OS cell lines (MG-63, U2-OS, and 143B) and a nude mice model generated by transplanting 143B cells. The cells and mice were treated with DMSO (control) or gradient concentrations of 3-AP. Then, various assays (e.g., cell counting kit-8, flow cytometry, immunohistochemistry, and western blotting) were performed to assess cell viability and apoptosis levels, as well as γH2A.X (DNA damage correlation), ribonucleotide reductase catalytic subunit M1 and M2 (RRM1 and RRM2, respectively) protein levels (iron-dependent correlation). 3-AP time- and dose-dependably suppressed growth and induced apoptosis in all three OS cell lines, and ferric ammonium citrate (FAC) blocked these effects. Moreover, 3-AP decreased RRM2 and total ribonucleotide reductase (RRM1 plus RRM2) protein expression but significantly increased γH2A.X expression; treatment did not affect RRM1 expression. Again, FAC treatment attenuated these effects. In vivo, the number of apoptotic cells in the tumor slices increased in the 3-AP-treated mice compared to the control mice. 3-AP treatment also decreased Ki-67 and p21 expression, suggesting inhibited OS growth. Furthermore, the expression of RRM1, RRM2, and transferrin receptor protein 1 (i.e., Tfr1) indicated that 3-AP inhibited OS growth via an iron-dependent pathway. In conclusion, 3-AP exhibits anticancer activity in OS by decreasing the activity of iron-dependent pathways, which could be a promising therapeutic strategy for OS.© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.