The Anti-tumor Activity of Flavokawain B and Zerumbone on Human Lung Carcinoma A549 Cells

• Main Authors: Yu-Chi Huang, 黃鈺琪
• Other Authors: 楊新玲
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/3wtmzj

碩士 === 中國醫藥大學 === 營養學系碩士班 === 102 === According to statistics, lung cancer is the leading cause of cancer deaths worldwide, it has 80 % of cancer patients pass away from lung cancer. Lung cancer is poor to diagnose, because of a lack of metastatic tumors and systemic treatment. Kava lactone is mainly extracts of kava (kava; Piper methysticum). Kava extract is known in the South Pacific islands that can be used to reduce the incidence of cancer. Kava chalcone include Flavokawain A、Flavokawain B and Flavokawain C, which have anti -tumor activity of different levels.Especially Flavokawain B. The results show that Flavokawain B IC50 =10μg/mL, and found as the dose ROS production.Autophagy also with the dose-dependent increased. As indicated by an increase in microtubule-associated protein light chain-3 puncta, accumulation of LC3II, and elevation of autophagic flux. The AVO assay also can observed the formation of autophagic vesicles. In addition , with the increase Flavokawain B concentrations , apoptosis index protein such as Bax、 caspase 3 and caspase 9 were increased , Bcl-2 was decreased. TUNEL assay also confirmed on apoptosis circumstances of A549 cells caused by the high concentration Flavokawain B. Recent studies have found that autophagy has two roles in the cell death process, cell survival or cell death. In our experiment, the autophagy inhibitor Bafilomycin A1, cell viability decreased. 3-MA and Chloroquine made cell viability rise that knows autophagy plays a role in cell death at Flavokawain B made A549 apoptosis. Therefore, Flavokawain B should be considered in the clinical application of lung cancer cells induced autophagy role as information assisted treatment of lung cancer. In the second part of the experiment, the literature indicates that TGF-β can promote epithelial- mesenchymal transition function may contribute to lung cancer metastasis. Zingiber Zerumbet Smith / Zingiber aromaticum major extract—Zerumbone, investigating the efficacy of inhibiting cancer metastasis of lung cancer when stimulated by the TGF-β. The results showed that TGF-β can induce EMT on A549 lung cancer cells and the cells were spindle-shaped morphology change into fusiform morphology. It also promotes cell migration and invasion effect.However, given Zerumbone at 10 and 20 μM can inhibit cancer cells and inhibition of MET in lung cancer cell migration and invasion. Western blot assay showed that EMT indicators protein E-cadherin gives a significant reduction in TGF-β stimulation. After treating Zerumbone, E-cadherin was returned and vimentin was unchanged.It also have the same result in the immunofluorescence assay. In addition,we found that giving Zerumbone could observed Smad2 phosphorylation was inhibited.In summary, TGF-β1-induced EMT and acetylation of Smad2 is negatively regulated by Zerumbone in non-small cell lung cancer.