Department of Traditional Chinese Medicine, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127, P.R. China.
Oncol Rep. 2014 Jan;31(1):305-13. doi: 10.3892/or.2013.2815. Epub 2013 Oct 24.
Ginsenosides, extracted from the traditional Chinese herb ginseng, are a series of novel natural anticancer products known for their favorable safety and efficacy profiles. The present study aimed to investigate the cytotoxicity of ginsenoside Rf to human osteosarcoma cells and to explore the anticancer molecular mechanisms of ginsenoside Rf. Five human osteosarcoma cell lines (MG-63, OS732, U-2OS, HOS and SAOS-2) were employed to investigate the cytotoxicity of ginsenoside Rf by MTT and colony forming assays. After treatment with ginsenoside Rf, MG-63 cells which were the most sensitive to ginsenoside Rf, were subjected to flow cytometry to detect cell cycle distribution and apoptosis, and nuclear morphological changes were visualized by Hoechst 33258 staining. Caspase-3, -8 and -9 activities were also evaluated. The expression of cell cycle markers including cyclin B1 and Cdk1 was detected by RT-PCR and western blotting. The expression of apoptotic genes Bcl-2 and Bax and the release of cytochrome c were also examined by western blotting. Change in the mitochondrial membrane potential was observed by JC-1 staining in situ. Our results demonstrated that the cytotoxicity of ginsenoside Rf to these human osteosarcoma cell lines was dose-dependent, and the MG-63 cells were the most sensitive to exposure to ginsenoside Rf. Additionally, ginsenoside Rf induced G2/M phase cell cycle arrest and apoptosis in MG-63 cells. Furthermore, we observed upregulation of Bax and downregulation of Bcl-2, Cdk1 and cyclin B1, the activation of caspase-3 and -9 and the release of cytochrome c in MG-63 cells following treatment with ginsenoside Rf. Our findings demonstrated that ginsenoside Rf induces G2/M phase cell cycle arrest and apoptosis in human osteosarcoma MG-63 cells through the mitochondrial pathway, suggesting that ginsenoside Rf, as an effective natural product, may have a therapeutic effect on human osteosarcoma.
人参皂苷是从传统中药人参中提取的一系列新型天然抗癌产品，以其良好的安全性和疗效而闻名。本研究旨在探讨人参皂苷 Rf 对人骨肉瘤细胞的细胞毒性，并探讨人参皂苷 Rf 的抗癌分子机制。本研究采用 MTT 和集落形成实验研究了人参皂苷 Rf 对 5 个人骨肉瘤细胞系（MG-63、OS732、U-2OS、HOS 和 SAOS-2）的细胞毒性。用流式细胞术检测经人参皂苷 Rf 处理后对人骨肉瘤细胞 MG-63 细胞周期分布和凋亡的影响，并用 Hoechst 33258 染色观察核形态变化。还评估了 caspase-3、-8 和 -9 的活性。通过 RT-PCR 和 Western blot 检测细胞周期标志物 cyclin B1 和 Cdk1 的表达。通过 Western blot 检测凋亡基因 Bcl-2 和 Bax 的表达以及细胞色素 c 的释放。通过 JC-1 染色原位观察线粒体膜电位的变化。结果表明，人参皂苷 Rf 对这些人骨肉瘤细胞系的细胞毒性呈剂量依赖性，MG-63 细胞对人参皂苷 Rf 的敏感性最高。此外，人参皂苷 Rf 诱导 MG-63 细胞 G2/M 期细胞周期阻滞和凋亡。此外，我们观察到在 MG-63 细胞中，人参皂苷 Rf 处理后 Bax 上调，Bcl-2、Cdk1 和 cyclin B1 下调，caspase-3 和 -9 激活以及细胞色素 c 释放。这些结果表明，人参皂苷 Rf 通过线粒体途径诱导人骨肉瘤 MG-63 细胞 G2/M 期细胞周期阻滞和凋亡，提示人参皂苷 Rf 作为一种有效的天然产物，可能对人骨肉瘤具有治疗作用。
