龍葵多酚抑制黑色素瘤細胞轉移及血管新生之作用與機轉

中山醫學大學機構典藏 CSMUIR > 醫學院 > 生化微生物免疫研究所 > 博碩士論文 > Item 310902500/436

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题名:	龍葵多酚抑制黑色素瘤細胞轉移及血管新生之作用與機轉 Chemoinhibitory Effect of Solanum Nigrum L. Polyphenol Extract on the Metastasis and Angiogenesis in B16 Melanoma Cells
作者:	吳敦豪 Dun-Hao Wu
贡献者:	中山醫學大學：生化暨生物科技研究所王朝鐘
关键词:	黑色素瘤轉移 PI3K/Akt NF-κB PKC 龍葵 melanoma migration metastasis Solanum nigrum L.
日期:	2009/07/23
上传时间:	2010-01-25T07:05:54Z (UTC)
摘要:	龍葵，為茄科植物分布於全球的耕地。在民間用於清熱解毒消腫，在醫學功能上已被證實具有抗氧化、抗發炎、護肝作用之功效。本篇研究目的是觀察龍葵水萃物(SNWE) 及龍葵多酚萃取物(SNPE) 是否能夠抑制腫瘤的轉移及與其密不可分的血管新生作用。首先，利用細胞毒性實驗、傷口癒合、移動分析、MMP電泳實驗來觀察SNWE 及SNPE 是否可以在in vitro 抑制B16-F1細胞轉移，結果顯示在不殺死細胞的前提下處理SNWE 及SNPE 後，細胞傷口癒合及移動能力明顯變差，而MMP-2及MMP-9的表現量亦是降低。進一步以西方墨點法分析與轉移相關的蛋白，實驗數據顯示Ras、PI3K/Akt、NF-κB、PKCα、PKCδ、JNK-p、P38-p 蛋白表現量皆明顯下降。此外，與細胞骨架有關的蛋白RhoA、CDC42，在處理SNWE 後其表現量也隨之降低。接著，以動物模式驗證龍葵抑制腫瘤轉移的作用，利用尾靜脈注射的方式將B16-F1細胞打入鼠體內並同時餵食SNWE，發現腫瘤轉移到肺臟或其他臟器的現象有得到改善，而腫瘤的生長也明顯受到抑制。另外，透過雞胚胎實驗及Plug assay，證實SNWE 及SNPE 皆能抑制血管新生作用。綜合以上的結果，推測龍葵具有降低腫瘤轉移及抗血管新生作用的潛力。 Solanum nigrum L., which belongs to the Solanaceae family, has been used traditionally to treat various ailment such as pain, inflammation and fever. It is believed to have anti-oxidative, anti-inflammatory, hepatoprotection. The purpose of this study were to investigate the inhibitory effects of water extract of Solanum nigrum L. (SNWE) and polyphenolic extract of Solanum nigrum L. (SNPE) on the angiogenesis and metastasis in B16-F1 cells. First, SNWE and SNPE exhibited an inhibitory effect on the migration ability by wound healing assay and Boyden chamber assay. Both SNWE and SNPE showed to reduce MMP-2 and MMP-9 in B16-F1 cells. The results of the Western blotting assay demonstrated that the expression levels of Ras, PI3K, phospho-Akt, and NF-κB in the SNWE and SNPE treated B16-F1 cells were reduced ; the expression levels of PKCα, PKCδ, phospho-JNK, phospho-p38 were also decreased, and such as phospho-ERK was unaffected. In angiogenesis experiment, SNWE and SNPE exhibited an inhibitory effect on the angiogenesis by chorioallantonic membrane inoculation and plug assay. Further investigation revealed that the anti-metastatic effect of SNWE was also evident in a C57Bl/6 mice model. The effects of dietary administration of SNWE on metastasis induced by B16-F1 cell in C57BL/6 mice. Accompanied with treated with SNWE at high concentrations indicated mice viability better. Taken together, the findings proved the inhibitory effect of SNWE and SNPE on the angiogenesis and metastasis of melanoma cells in vitro and in vivo.
URI:	http://140.128.138.153:8080/handle/310902500/436
显示于类别:	[生化微生物免疫研究所] 博碩士論文

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