在細胞及動物模式探討洛神花青素抑制轉移及血管新生之作用

洛神花為錦葵科的成員，主要活性成分含有兒茶酸，花青素，類黃酮和異黃酮，傳統的醫學上洛神花具清熱、解渴、止咳、降血壓之功效。轉移和血管新生息息相關，是在癌症的晚期的主要致死原因。阻止或延緩腫瘤的轉移和血管新生將是一個潛在的治療方法。本篇是觀察洛神分離出來的花青素（Hibiscus Anthocyanins）抑制腫瘤轉移和血管新生之作用。首先，MTT分析HAs對B16-F1細胞之生長作用，發現其 IC50為 2.46 mg/ml，後續選擇此濃度進行實驗，進行Wound healing和Gelatin-zymography，分析在細胞抗腫瘤轉移之作用，結果顯示，隨著劑量增加B16-F1細胞的轉移之能力顯著的被抑制。另外，用HUVEC細胞進行tube formation assay探討HAs對血管新生作用的影響，結果顯示，HAs有效降低HUVEC血管新生之作用。接著，運用Western blot探討在B16-F1和HUVEC細胞中HAs抑制腫瘤轉移及血管新生之機制，發現HAs藉由抑制VEGF/VEGF-R及其下游Ras 、PI3K/Akt、PKC及MAPK達到抑制轉移和血管新生之作用。進一步，利用C57BL / 6小鼠囀移動物模式和裸鼠plus assay分析，探討生物體中HAs的處理抑制腫瘤轉移和血管新生。結果顯示，HAs在生物體內可以抑制腫瘤轉移及血管新生。綜上所述，我們發現無論細胞實驗或動物模式，HAs對於抑制腫瘤的轉移和血管新生具有良好的功效。希望在這項研究的初步結果可以應用在未來的藥物開發和輔助治療保健食品的發展。

關鍵字

黑色素瘤；轉移；血管新生；洛神

並列摘要

Roselle is a member of the Malvaceae family the main active ingredient contains catechins, anthocyanins, flavonoids and isoflavones. As a folk medicine, Roselle can cure high blood pressure, heat stroke, drunk, cough. Metastasis and angiogenesis occurs at the later stage of cancer, and also is the main cause of death. The potential therapeutic strategy to combat this devastating disease is to delay or prevent tumor metastasis and angiogenesis. Previous evidence supports that Hibiscus anthocyanins (HAs) reduce tumorigenesis, but the mechanisms by which these HAs inhibit metastasis and angiogenesis are poorly understood. MTT was used to analyze the growth-inhibiting effect of HAs. HAs complied with a dose-dependent repression of cancer cell proliferation with an IC50 (50% inhibitory concentration) value of 2.46 mg/ml. The concentration of below 1 mg/ml is recommended to begin further studies. Here we showed that HAs inhibit migration of B16-F1 cells by wound healing and gelatin-zymography analysis. In addition, exposure of human umbilical endothelial cells (HUVECs) to 0.1 to 2.0 μM HAs significantly blocked tube formation. We investigate the detailed mechanism by which HAs inhibits angiogenic effects in B16 and HUVECs cells using Western blot assay. Under the same concentrations, HAs markedly decreased VEGF-stimulated activation of VEGF receptor, and its downstream Ras, PI3K/Akt, PKC as well as mitogen-activated protein kinases. Furthermore, HAs exhibited an inhibitory effect on migration and angiogenesis by C57BL/6 mice and plus assay. Taken together, the findings proved the inhibitory effect of HAs on tumor metastasis and angiogenesis in vitro and in vivo. Hope that the preliminary results of this study can be applied to future drug development and the health food development.

並列關鍵字

Melanoma ； metastasis ； angiogenesis ； Roselle

參考文獻

1. Chang, Y.C., et al., Hibiscus anthocyanins-rich extract inhibited LDL oxidation and oxLDL-mediated macrophages apoptosis. Food Chem Toxicol, 2006. 44(7).

2. Palfi, A., et al., Alcohol-free red wine inhibits isoproterenol-induced cardiac remodeling in rats by the regulation of Akt1 and protein kinase C alpha/beta II. J Nutr Biochem, 2009. 20(6).

3. Hamalainen, M., et al., Anti-inflammatory effects of flavonoids: genistein, kaempferol, quercetin, and daidzein inhibit STAT-1 and NF-kappaB activations, whereas flavone, isorhamnetin, naringenin, and pelargonidin inhibit only NF-kappaB activation along with their inhibitory effect on iNOS expression and NO production in activated macrophages. Mediators Inflamm, 2007.

4. Hirose, Y., et al., Chemoprevention of urinary bladder carcinogenesis by the natural phenolic compound protocatechuic acid in rats. Carcinogenesis, 1995. 16(10).

5. Tseng, T.H., et al., Induction of apoptosis by hibiscus protocatechuic acid in human leukemia cells via reduction of retinoblastoma (RB) phosphorylation and Bcl-2 expression. Biochem Pharmacol, 2000. 60(3).

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在細胞及動物模式探討洛神花青素抑制轉移及血管新生之作用

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