3-氨基吡啶-2-甲醛缩氨基硫脲(3-AP)具有广谱抗肿瘤活性。然而,其在骨肉瘤(OS)中的作用尚不清楚。因此,本研究使用三种人OS细胞系(MG-63,U2-OS,和143B)和通过移植143B细胞产生的裸鼠模型。用DMSO(对照)或梯度浓度的3-AP处理细胞和小鼠。然后,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)进行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)蛋白水平(铁依赖性相关性)。3-AP在所有三种OS细胞系中时间和剂量依赖性地抑制生长并诱导凋亡,柠檬酸铁铵(FAC)阻断了这些影响。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表达;处置不影响RRM1表达。再一次,FAC治疗减弱了这些作用。在体内,与对照小鼠相比,在3-AP处理的小鼠中肿瘤切片中的凋亡细胞的数量增加。3-AP治疗也降低了Ki-67和p21的表达,表明OS增长受到抑制。此外,RRM1、RRM2和转铁蛋白受体蛋白1的表达(即,Tfr1)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而在OS中表现出抗癌活性,这可能是一种有前途的OS治疗策略。
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.