2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38079Cancer prevention has been a world trend. Because of this, searching for chemopreventive agents is a very important thing. Anthocyanins and polyphenol are abundant in vegetables and fruits. Many reports indicated they have function in anti-oxidation, reducing cholesterol, and preventing cancers. Our laboratory explores anthocyanins extracted from Hibiscus sabdariffa Linne, Hibiscus anthocyanins (HAs), have antioxidant activity and liver protection (Wang et al., Fd. Chem. Toxicol. 35, 1159-1164, 1997; 38, 411-416, 2000). Recent data also shown that HAs and HPE promote AGS, 3B, G2, KB, HL-60, MCF-7, Caco-2 cells apoptosis. Thus, this research investigated that HAs, HPE anticancer function and its molecular mechanism. The first year, we analyzed HSE, HAs function and its functional component HAs and HPE work in anticancer. The working items: (1) using flow cytometery and DNA electrophoresis analyzed HSE, HAs and HPE promote all kinds of human cancer cells apoptosis; (2) exploring apoptosis-related gene expression, including bcl-2 family, caspase, cytochrome c et al.; (3) testing HSE, HAs and HPE affect cancer cells in cell cycle, including several cyclins, CDKs and their association, Rb phosphorylation, with E2F association, cell cycle distribution. Take together, we expect that HSE, HAs and HPE promote cancer cell apoptosis and inhibit cell cycle progression. The mechanisms were through activation of p38/FasL and p53/Bax cascade (supplements paper 1 and 2). Based on we found HPE and HAs have the best effect to AGS, HL-60 cells, respectively, hence, this year work has used two animal models to study: benzo(a)pyrene-induced gastric cancer of Swiss albino mice and N-Nitroso-N-Methylurea (NMU)-induced leukemia of Sprague-Dawley rat. In addition, our laboratory have data shown that HPE and HAs could affected the expression of MMP, therefore, next year, experiments are design to explored HPE and HAs as metastasis inhibitor evaluated by migration, adhesion and invasion assay and exhibits a key role in inhibition of metastasis-associated metalloproteinase secretion. To investigate whether HPE and HAs could also affect tumor metastasis in vivo, we design an animal model for the inhibition of lung metastasis induced by B16 melanoma cells in mice. According to these studies, it would demonstrate HSE and its compound HPE, HAs have the effect of anticancer and cancer prevention. Furthermore, we apply IND to progress human clinical test of traditional herb. We expect it could application for developing a healthy food and a new chemopreventive agent.癌症預防已為一世界潮流，因此，尋找一化學預防物質（chemopreventive agent）為一重要工作。花青素（Anthocyanins）及多酚（Polyphenol）廣存於蔬菜、水果等植物中，已有許多報告證明花青素及多酚具抗氧化活性，降低血脂肪及防癌等作用，本研究室也發現分離自洛神花之花青素（Hibiscus anthocyanins, HAs）有抗氧化及保肝之功能（Wang et al., Fd. Chem. Toxicol. 35, 1159-1164, 1997；38, 411-416, 2000）。最近本實驗室的研究顯示此類HAs及Hibiscus polyphenol（HPE）會促進胃癌（AGS），肝癌（3B, G2），口腔上皮癌（KB），血癌（HL-60），乳癌（MCF-7），腸癌（Caco-2）等細胞凋謝死亡，因此，本研究擬深入探討HAs, HPE之抗癌作用及其作用之分子機制。 上一年度的工作探討洛神花萃取物（Hibiscus Sabdaffra extracts, HSE）及其功能性成份HAs及HPE抗癌之作用，包括（1）利用flow cytometer 及DNA electrophoresis 分析HSE, HAs, HPE促進各種癌細胞凋謝死亡；（2）探討apoptosis-related gene expression，包括bcl-2 family, caspase, cyt c；（3）探討HSE, HAs, HPE對癌細胞cell cycle的影響，包括各種Cyclins及CDKs及它們的association, Rb phosphorylation及與E2F之association, cell cycle分佈等，由以上的結果我們發現HSE, HAs, HPE可促進癌細胞凋謝死亡及抑制cell cycle progression，其機轉為活化p38/FasL cascade及p53/Bax路徑(附件一, 二)。由於HPE對AGS(胃癌細胞)及HAs對HL-60(血癌細胞)之作用最敏感，因此，本年度我們將利用benzo(a)pyrene致小白鼠(Swiss albino mice)胃癌及N-Nitroso-N-Methylurea (NMU)致雄性大白鼠（Sprague-Dawley rat）血癌之模式，且分別進行HSE抑制老鼠胃癌及血癌之作用。另外，本實驗室初步證據顯示，HPE及HAs影響MMP之表現，因此，下一年的工作，我們將研究HPE及HAs抑制癌細胞的轉移，擬利用migration assay, attachment assay及invasion assay，並使用semi-RT-PCR及gelatin zymography assay探討metalloproteinase activity，並進一步以小鼠體內之B16細胞轉移模式觀察抑制癌細胞metastasis之作用，以及深入探討其抑制惡化轉移之機制。 從以上細胞及動物之研究，我們將獲得HSE及其成分HPE與HAs之防癌、抗癌作用，並發展為保健食品，進一步申請IND執行中草藥人體臨床試驗，發展為抗癌新藥。花青素洛神多酚胃癌血癌金屬蛋白惡化轉移動物模式AnthocyaninsPolyphenolgastric cancerLeukemiametalloproteinasemetastasisanimal modelProject