Department of Orthopaedics, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
J Zhejiang Univ Sci B. 2011 Apr;12(4):247-55. doi: 10.1631/jzus.B1000373.
This paper aims to investigate the effects of artesunate (ART) on growth and apoptosis in human osteosarcoma HOS cell line in vitro and in vivo and to explore the possible underlying mechanisms. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The induction of apoptosis was detected by light and transmission electron microscopy and flow cytometry. Western blot analysis was used to investigate the related mechanisms. Nude mice were further employed to investigate the antitumour activity of ART in vivo. MTT assay results demonstrated that ART selectively inhibits the growth of HOS cells in a dose- and time-dependent manner. Based on the findings of light and transmission electron microscopy, Hoechst 33258 staining, and fluorescein isothiocyanate (FITC)-annexin V staining, the cytotoxicity of ART in HOS cells occurs through apoptosis. With ART treatment, cytosolic cytochrome c was increased, Bax expression was gradually upregulated, Bcl-2 expression was downregulated, and caspase-9 and caspase-3 were activated. Thus, the intrinsic apoptotic pathway may be involved in ART-induced apoptosis. Cell cycle analysis by flow cytometry indicated that ART may induce cell cycle arrest at G(2)/M phase. In nude mice bearing HOS xenograft tumours, ART inhibited tumour growth and regulated the expressions of cleaved caspase-3 and survivin, in agreement with in vitro observations. ART has a selective antitumour activity against human osteosarcoma HOS cells, which may be related to its effects on induction of apoptosis via the intrinsic pathway. The results suggest that ART is a promising candidate for the treatment of osteosarcoma.
本文旨在研究青蒿琥酯（ART）在体外和体内对人骨肉瘤 HOS 细胞系生长和凋亡的影响，并探讨其可能的作用机制。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）比色法检测细胞活力。通过光镜和透射电镜以及流式细胞术检测细胞凋亡的诱导。采用 Western blot 分析探讨相关机制。进一步采用裸鼠研究 ART 在体内的抗肿瘤活性。MTT 比色法结果表明，ART 呈剂量和时间依赖性选择性抑制 HOS 细胞生长。基于光镜和透射电镜、Hoechst 33258 染色和异硫氰酸荧光素（FITC）- Annexin V 染色的结果，ART 在 HOS 细胞中的细胞毒性通过凋亡发生。随着 ART 处理，细胞质细胞色素 c 增加，Bax 表达逐渐上调，Bcl-2 表达下调，caspase-9 和 caspase-3 被激活。因此，内在凋亡途径可能参与了 ART 诱导的凋亡。流式细胞术的细胞周期分析表明，ART 可能诱导细胞周期停滞在 G(2)/M 期。在荷人骨肉瘤 HOS 移植瘤裸鼠中，ART 抑制肿瘤生长，并调节 cleaved caspase-3 和 survivin 的表达，与体外观察结果一致。ART 对人骨肉瘤 HOS 细胞具有选择性抗肿瘤活性，这可能与其通过内在途径诱导细胞凋亡有关。结果表明，ART 是治疗骨肉瘤的一种有前途的候选药物。
