Lee Hwa-Jeong, Son Dae-Hyung, Lee Sun-Kyung, Lee Jun, Jun Chang-Duk, Jeon Byung-Hun, Lee Suk-Keun, Kim Eun-Cheol
Department of Oral and Maxillofacial Pathology, College of Dentistry, Wonkwang University, Iksan, South Korea.
Phytother Res. 2006 Sep;20(9):773-9. doi: 10.1002/ptr.1956.
Coptidis rhizoma (C. rhizoma) had been demonstrated as an antioxidant and anticancer agent, however, its antioral cancer mechanism still remains unclear. Using water extracts of C. rhizoma, growth and apoptosis-related experiments for the treatment of multi-stage of oral cancer were carried out on immortalized human oral keratinocytes (IHOK), primary oral cancer cells (HN4), metastatic oral cancer cells (HN12) and human skin keratinocytes (HaCaT) by MTT assay, three-dimensional (3-D) raft cultures, western blotting, cell cycle analysis, nuclear staining and cytochrome c expression related to the apoptosis signaling pathway. C. rhizoma inhibited the proliferation of immortalized and malignant oral keratinocytes in a dose- and time-dependent manner. In 3-D organotypic culture, C. rhizoma-treated cells showed less maturation than the control cells, displaying low surface keratinization and decreased epithelial thickness. The major mechanism of growth inhibition by C. rhizoma appears to be the induction of apoptosis, which is supported by the results of the cell cycle analysis, FITC-annexin V staining, DNA fragmentation assay and DAPI staining. The induction of apoptosis by C. rhizoma was more prominent in immortalized keratinocytes than in malignant oral keratinocytes. Cytochrome-c release from mitochondria, accompanied by the activation of caspase-3, was observed in C. rhizoma-treated IHOK and oral cancer cells. These results suggest that C. rhizoma has apoptotic effects in immortalized and malignant oral keratinocytes via the mitochondrial signaling pathway.
黄连已被证明是一种抗氧化剂和抗癌剂，然而，其抗口腔癌的机制仍不清楚。利用黄连水提取物，通过MTT法、三维（3-D）筏培养、蛋白质免疫印迹法、细胞周期分析、核染色以及与凋亡信号通路相关的细胞色素c表达，对永生化人口腔角质形成细胞（IHOK）、原发性口腔癌细胞（HN4）、转移性口腔癌细胞（HN12）和人皮肤角质形成细胞（HaCaT）进行了多阶段口腔癌治疗的生长和凋亡相关实验。黄连以剂量和时间依赖性方式抑制永生化和恶性口腔角质形成细胞的增殖。在三维器官型培养中，黄连处理的细胞比对照细胞成熟度更低，表现为表面角质化程度低和上皮厚度降低。黄连抑制生长的主要机制似乎是诱导凋亡，这得到了细胞周期分析、FITC-膜联蛋白V染色、DNA片段化分析和DAPI染色结果的支持。黄连诱导的凋亡在永生化角质形成细胞中比在恶性口腔角质形成细胞中更显著。在黄连处理的IHOK和口腔癌细胞中观察到线粒体释放细胞色素c，并伴有caspase-3的激活。这些结果表明，黄连通过线粒体信号通路在永生化和恶性口腔角质形成细胞中具有凋亡作用。
