Tang Yih-Jing, Yang Jai-Sing, Lin Chin-Fen, Shyu Woei-Cherng, Tsuzuki Minoru, Lu Chi-Cheng, Chen Yuh-Fung, Lai Kuang-Chi
Department of Family Medicine, Taichung Veterans General Hospital, School of Medicine, Chung-Shan Medical University, Taichung, Taiwan, ROC.
Oncol Rep. 2009 Nov;22(5):1051-6. doi: 10.3892/or_00000535.
The Houttuynia cordata Thunb (HCT) extract has been used as a traditional Chinese herb medicine and as well as an effective drug for treating allergic inflammation for thousands of years. In this study, we investigated the anti-cancer activity of HCT and its molecular mechanisms in the human colon adenocarcinoma cell line HT-29. HCT inhibited HT-29 cell viability in a dose- and time-dependent manner by MTT assay. Treatment with 450 microg/ml of HCT for 48 and 72 h led to DNA damage and apoptosis by DAPI staining and comet assay. HCT increased reactive oxygen species production and decreased the levels of mitochondria membrane potential (MMP) in HT-29 cells by flow cytometry analysis. HCT caused the release of cytochrome c, Apaf-1, pro-caspase-9 and AIF from mitochondria via a decrease of the MMP. The decrease of MMP was then associated with a decrease in the ratio of Bax/Bcl-2 and activation of caspase-9 and -3 by Western blotting and caspase activity assay. Caspase-9 and -3 inhibitors almost completely suppressed HCT-induced caspase-9 and -3 activities. Our results demonstrated that the HCT-induced apoptosis in human colon adenocarcinoma cell line HT-29 might be related to a mitochondrial-dependent pathway.
鱼腥草提取物作为一种传统中药，数千年来一直被用作治疗过敏性炎症的有效药物。在本研究中，我们研究了鱼腥草在人结肠腺癌细胞系HT - 29中的抗癌活性及其分子机制。通过MTT法检测，鱼腥草以剂量和时间依赖性方式抑制HT - 29细胞活力。用450μg/ml鱼腥草处理48小时和72小时，通过DAPI染色和彗星试验导致DNA损伤和细胞凋亡。通过流式细胞术分析，鱼腥草增加了HT - 29细胞中活性氧的产生，并降低了线粒体膜电位（MMP）水平。鱼腥草通过降低MMP导致细胞色素c、凋亡蛋白酶激活因子-1（Apaf - 1）、前半胱天冬酶-9和凋亡诱导因子从线粒体释放。然后，通过蛋白质免疫印迹法和半胱天冬酶活性测定，MMP的降低与Bax/Bcl - 2比值的降低以及半胱天冬酶-9和-3的激活相关。半胱天冬酶-9和-3抑制剂几乎完全抑制了鱼腥草诱导的半胱天冬酶-9和-3活性。我们的结果表明，鱼腥草诱导人结肠腺癌细胞系HT - 29凋亡可能与线粒体依赖性途径有关。
