* Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
† Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
Am J Chin Med. 2019;47(3):635-656. doi: 10.1142/S0192415X19500332. Epub 2019 Apr 25.
Cardamonin, the chalcone class, is one of the natural components from the spicy herbaceous plant ( Griff) and has anticancer activities in many human cancer cell lines. There is, however, no information to show that cardamonin induces cell apoptosis and alters apoptosis associated gene expressions in mouse leukemia cells. Thus, we investigated the effects of cardamonin on the apoptotic cell death and associated gene expression in mouse leukemia WEHI-3 cells . Results indicated that cardamonin decreased total viable cell number induced cell morphological changes and apoptotic cell death in WEHI-3 cells that were assay by contrast-phase microscopy and flow cytometry examinations, respectively. The flow cytometry assay indicated that cardamonin increased reactive oxygen species (ROS) and Ca production, decreased the levels of mitochondrial membrane potential ( and increased caspase-3, -8 and -9 activities in WEHI-3 cells. Western blotting was performed to analyze expression of relevant pro- and anti-apoptotic proteins and results showed that cardamonin decreased anti-apoptotic protein of Bcl-2 but increased pro-apoptotic protein of Bax in WEHI-3 cells. Furthermore, cardamonin increased cytochrome c, AIF and Endo G release, increased GRP78, caspase-12 that were associated with ER stress and increased Fas, Fas-Ligand and FADD expression. Furthermore, cardamonin increased the gene expressions of (death-associated protein), transmembrane (BAX inhibitor motif containing 4), (autophagy related 5) but decreased the gene expression of (DNA-damage inducible transcript 3), (DNA-damage-inducible transcript 4), (BCL2-associated athanogene 6), [BCL2-like 13 (apoptosis facilitator)] and (BRCA1-associated ATM activator 1) that are associated with apoptosis pathways. Based on those findings, we may suggest cardamonin induced apoptotic cell death through Fas and Fas-Ligand-, caspase- and mitochondria-dependently pathways and also affects the apoptotic gene expression in WEHI-3 cells .
小豆蔻明，查尔酮类，是从辛辣草本植物（ Griff ）中的一种天然成分，具有多种人类癌细胞系的抗癌活性。然而，没有信息表明小豆蔻明诱导细胞凋亡并改变小鼠白血病细胞中与凋亡相关的基因表达。因此，我们研究了小豆蔻明对小鼠白血病 WEHI-3 细胞凋亡细胞死亡和相关基因表达的影响。结果表明，小豆蔻明降低总活细胞数，通过相差显微镜和流式细胞术分别观察到细胞形态变化和 WEHI-3 细胞的凋亡细胞死亡。流式细胞术检测表明，小豆蔻明增加活性氧（ROS）和 Ca2+产生，降低线粒体膜电位（Δψm）水平，并增加 WEHI-3 细胞中 caspase-3、-8 和 -9 的活性。进行 Western blot 分析以分析相关促凋亡和抗凋亡蛋白的表达，结果表明小豆蔻明降低了 WEHI-3 细胞中的抗凋亡蛋白 Bcl-2，但增加了促凋亡蛋白 Bax。此外，小豆蔻明增加了细胞色素 c、AIF 和 Endo G 的释放，增加了与内质网应激相关的 GRP78、caspase-12，并增加了 Fas、Fas-Ligand 和 FADD 的表达。此外，小豆蔻明增加了（死亡相关蛋白）、（跨膜（含有 BAX 抑制剂基序 4））、（自噬相关 5）的基因表达，但降低了（DNA 损伤诱导转录 3）、（DNA 损伤诱导转录 4）、（BCL2 相关的athanogene 6）、（BCL2 样 13（凋亡促进因子））和（BRCA1 相关的 ATM 激活剂 1）的基因表达，这些基因与凋亡途径有关。基于这些发现，我们可以提出小豆蔻明通过 Fas 和 Fas-Ligand-、caspase 和线粒体依赖性途径诱导凋亡细胞死亡，并且还影响 WEHI-3 细胞中的凋亡基因表达。
