School of Medicine, Chung Shan Medical University, Taichung 402, Taiwan.
Department of Orthopedic Surgery, Chung Shan Medical University, Taichung 402, Taiwan.
Am J Chin Med. 2015;43(5):1031-42. doi: 10.1142/S0192415X15500597. Epub 2015 Jul 30.
In the past decade, no significant improvement has been made in chemotherapy for osteosarcoma (OS). To develop improved agents against OS, we screened 70 species of medicinal plants and treated two human OS cell lines with different agent concentrations. We then examined cell viability using the MTT assay. Results showed that a candidate plant, particularly the rhizomes of Anemone altaica Fisch. ex C. A. Mey aqueous extract (AAE), suppressed the viability of HOS and U2OS cells in a concentration-dependent manner. Flow cytometry analysis revealed that AAE significantly increased the amount of cell shrinkage (Sub-G1 fragments) in HOS and U2OS cells. Moreover, AAE increased cytosolic cytochrome c and Bax, but decreased Bcl-2. The amount of cleaved caspase-3 and poly-(ADP-ribose) polymerase-1 (PARP-1) were significantly increased. AAE suppressed the growth of HOS and U2OS through the intrinsic apoptotic pathway. Data suggest that AAE is cytotoxic to HOS and U2OS cells and has no significant influence on human osteoblast hFOB cells. The high mRNA levels of apoptosis-related factors (PPP1R15A, SQSTM1, HSPA1B, and DDIT4) and cellular proliferation markers (SKA2 and BUB1B) were significantly altered by the AAE treatment of HOS and U2OS cells. Results show that the anticancer activity of AAE could up-regulate the expression of a cluster of genes, especially those in the apoptosis-related factor family and caspase family. Thus, AAE has great potential as a useful therapeutic drug for human OS.
在过去的十年中，骨肉瘤（OS）的化疗并没有取得显著进展。为了开发针对 OS 的改良药物，我们筛选了 70 种药用植物，并以不同浓度的药物处理两种人骨肉瘤细胞系。然后使用 MTT 测定法检查细胞活力。结果表明，候选植物，特别是阿尔泰银莲花（Anemone altaica Fisch. ex C. A. Mey）根茎的水提取物（AAE），以浓度依赖性方式抑制 HOS 和 U2OS 细胞的活力。流式细胞术分析显示，AAE 显着增加 HOS 和 U2OS 细胞中细胞皱缩（Sub-G1 片段）的量。此外，AAE 增加了细胞质细胞色素 c 和 Bax，但减少了 Bcl-2。 cleaved caspase-3 和多聚（ADP-核糖）聚合酶-1（PARP-1）的量显着增加。 AAE 通过内在凋亡途径抑制 HOS 和 U2OS 的生长。数据表明，AAE 对 HOS 和 U2OS 细胞具有细胞毒性，对人成骨细胞 hFOB 细胞没有明显影响。 AAE 处理 HOS 和 U2OS 细胞后，凋亡相关因子（PPP1R15A、SQSTM1、HSPA1B 和 DDIT4）和细胞增殖标记物（SKA2 和 BUB1B）的高 mRNA 水平显着改变。结果表明，AAE 的抗癌活性可以上调一组基因的表达，特别是凋亡相关因子家族和半胱天冬酶家族的基因。因此，AAE 作为治疗人类骨肉瘤的有用药物具有很大的潜力。
