Department of Natural Products Chemistry, School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin, China.
Department of Pharmacy, The First Hospital of Jilin University, Changchun, Jilin, China.
J Biochem Mol Toxicol. 2020 Dec;34(12):e22597. doi: 10.1002/jbt.22597. Epub 2020 Aug 6.
Ginsenoside Rh2 is a primary bioactive compound obtained from ginseng that indicated anticancer activities against several malignant tumors. However, previous studies have reported little about the inhibitory effect of Rh2 on osteosarcoma (OS). This study aims to explore whether Rh2 could exert anticancer effects in OS cells and further investigate the proliferation, migration, and apoptosis mechanisms induced by Rh2 in human OS U20S cell line. The viability of U20S cells was obtained by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Cell migration property was analyzed by wound-healing assay. Apoptosis was visualized using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL), 4',6-diamidino-2-phenylindole (DAPI), and annexin V/propidium iodide (PI) staining. Relative protein expressed was confirmed through Western blot analysis. Mitochondrial membrane potential was evaluated by JC-1 staining. In this study, we used broad-spectrum anticancer drug cisplatin (CP) as a positive control. The results indicated that Rh2 remarkably inhibited cell viability of U20S cells in a dose- and time-dependent manner, and suppressed migration. TUNEL, DAPI, annexin V/PI, and JC-1 assay suggested that Rh2 could induce cellular apoptosis. Rh2 could reduce the levels of Bcl-2, caspase 3, and caspase 9, and promote the expression level of Bax in U20S cells. Moreover, Rh2 could induce apoptosis by promoting mitogen-activated protein kinase (MAPK) signaling pathway and inhibit PI3K/Akt/mTOR and nuclear factor-κB (NF-κB) signaling pathway in U20S cells. These findings indicated that Rh2 has an anticancer effect on U20S cells by regulating MAPK, PI3K/Akt/mTOR, and NF-κB signaling pathway.
人参皂苷 Rh2 是一种从人参中提取的主要生物活性化合物，对多种恶性肿瘤具有抗癌活性。然而，以前的研究很少报道 Rh2 对骨肉瘤（OS）的抑制作用。本研究旨在探讨 Rh2 是否能在 OS 细胞中发挥抗癌作用，并进一步研究 Rh2 诱导人骨肉瘤 U20S 细胞系增殖、迁移和凋亡的机制。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐法获得 U20S 细胞活力。通过划痕愈合试验分析细胞迁移特性。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）、4',6-二脒基-2-苯基吲哚（DAPI）和膜联蛋白 V/碘化丙啶（PI）染色观察细胞凋亡。通过 Western blot 分析确认相对蛋白表达。通过 JC-1 染色评估线粒体膜电位。在本研究中，我们使用广谱抗癌药物顺铂（CP）作为阳性对照。结果表明，Rh2 以剂量和时间依赖的方式显著抑制 U20S 细胞活力，并抑制迁移。TUNEL、DAPI、膜联蛋白 V/PI 和 JC-1 检测表明 Rh2 可诱导细胞凋亡。Rh2 可降低 U20S 细胞中 Bcl-2、caspase 3 和 caspase 9 的水平，并促进 Bax 的表达水平。此外，Rh2 可通过促进丝裂原活化蛋白激酶（MAPK）信号通路并抑制 U20S 细胞中的 PI3K/Akt/mTOR 和核因子-κB（NF-κB）信号通路诱导细胞凋亡。这些发现表明，Rh2 通过调节 MAPK、PI3K/Akt/mTOR 和 NF-κB 信号通路对 U20S 细胞具有抗癌作用。
