| Summary: | 碩士 === 國立高雄大學 === 生命科學系碩士班 === 102 === Bulnesia sarmientoi tree, which mainly distributed in South America, is a slow-growing hardwood species. The traditional use of B. sarmientoi can eliminate wind, heat, and anti-toxic function, as well as to promote skin healing, appetizers, spleen function, and brain doping. Previous studies indicate that B. sarmientoi has anti-inflammatory and anti-cancer activities. The goal of this study is to investigate the anti-proliferation effects of B. sarmientoi supercritical fluid extract (BSE) against A549, H661, H1299 human lung cancer cells and normal embryonic lung fibroblast cells MRC-5 in vitro, and its necroptotic property. The experimental results showed that the IC50 are 43.7, 44.6 and 53.3 μg/ml, repectively, for A549, H661 and H1299 cells under 48 hr treatment of BSE. In comparison, the IC50 is 89.7 μg/ml for MRC-5 normal cells. This implies that BSE has the cytototic specificity on the cancer cells. Additionally, a positive correlation existed between the treatment time and the inhibition of cell growth, indicating that BSE is a long-duration type of tumor suppressor agent. The flow cytometric analysis indicated that BSE could arrest cancer cells in G2/M phase of the cell cycle. Furthermore, the necroptotic effect of BSE on A549 and H661 cells was confirmed by annexin V-FITC/PI double staining. BSE induced lung cancer cells to the necroptosis pathway was further verified by Western blot analysis, since the pro-necroptotic TNF-α, RIP-1 and RIP-3 proteins were down-regulated and the anti- necroptotic caspase-8 was up-regulated. From the assays of wound healing, migration and invasion, BSE was proved to have a significant inhibitory activity on the migration of lung cancer cells. The results of Western blot analysis showed that the BSE treatment down-regulated the expression of MMP-2 and MMP-9. Taken together, since BSE possesses significant cytotoxicity on A549 and H661 cells and inhibitory activity on the metastasis of lung cancer cells, it may serve as a potential target for the treatment of lung cancer.
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