School of Life Science and Technology, Wuhan Polytechnic University, Wuhan, 430023, China.
National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China.
Mol Biol Rep. 2022 Jun;49(6):4247-4255. doi: 10.1007/s11033-022-07259-7. Epub 2022 Feb 25.
Chikusetsusaponin IV and V (CsIV and CsV), two typical oleanolic acid saponins, are mainly derived from the rhizome of Panax japonicus C.A. Mey. To reveal the anti-cancer effect of CsIV and CsV on liver cancer cells, human hepatic cancer cell lines (HepG2) were exposed to these saponins, and various physiological responses of HepG2 were investigated.
HepG2 cells were treated with CsIV, CsV and 5-fluorouracil (5-FU). Cell proliferation was measured by CCK-8 assay. The cell cycle arrest, cell apoptosis and intracellular Ca levels were respectively identified by flow cytometry. The mitochondrial membrane potential was detected by fluorescence microscopy. And, the levels of apoptosis-related proteins were analyzed by western blotting. Both CsIV and CsV were demonstrated to inhibit cell viability, and induce cell cycle arrest and apoptosis of HepG2 in a dose-dependent manner. They also enhanced the intracellular Ca level and decreased the mitochondrial membrane potential in HepG2 cells. Furthermore, p53 and p21 were found up-regulated in HepG2 cells treated by CsIV and CsV. The apoptotic proteins, bax, cytochrome c, cleaved caspase-3/-9, were all found activated in HepG2 cells after CsIV and CsV treatment. The anti-apoptotic protein, bcl-2, was significantly down-regulated in all treated HepG2 cells.
Our data demonstrated that CsIV and CsV exerted significant cytotoxic effects on HepG2 cells without affecting normal liver cells. And, these chikusetsusaponins, especially for CsIV, showed a potent effect on promoting cell apoptosis in HepG2 cells, which was associated with the activation of p53-mediated apoptosis pathway.
齐墩果酸皂甙 IV 和 V（CsIV 和 CsV）是两种典型的齐墩果酸皂甙，主要来源于人参 Panax japonicus C.A. Mey 的根茎。为了揭示 CsIV 和 CsV 对肝癌细胞的抗癌作用，将这些皂甙暴露于人肝癌细胞系（HepG2）中，并研究了 HepG2 的各种生理反应。
用 CsIV、CsV 和 5-氟尿嘧啶（5-FU）处理 HepG2 细胞。用 CCK-8 法测定细胞增殖。用流式细胞术分别鉴定细胞周期停滞、细胞凋亡和细胞内 Ca 水平。用荧光显微镜检测线粒体膜电位。并用 Western blot 分析凋亡相关蛋白的水平。结果表明，CsIV 和 CsV 均能抑制 HepG2 细胞活力，并呈剂量依赖性诱导细胞周期停滞和凋亡。它们还增强了 HepG2 细胞内的 Ca 水平并降低了线粒体膜电位。此外，在 CsIV 和 CsV 处理的 HepG2 细胞中发现 p53 和 p21 上调。凋亡蛋白 bax、细胞色素 c、cleaved caspase-3/-9 在 CsIV 和 CsV 处理的 HepG2 细胞中均被发现被激活。所有用 CsIV 和 CsV 处理的 HepG2 细胞中抗凋亡蛋白 bcl-2 均显著下调。
我们的数据表明，CsIV 和 CsV 对 HepG2 细胞具有显著的细胞毒性作用，而对正常肝细胞没有影响。而且，这些齐墩果酸皂甙，特别是 CsIV，在 HepG2 细胞中表现出促进细胞凋亡的强大作用，这与 p53 介导的凋亡途径的激活有关。
