中國橄欖萃取物及其主要成分對大腸直腸癌細胞的抑制功效

The inhibitory effect Chinese olive (Canarium album L.) extract and its major components on human colorectal cancer cells

龍孟專(Meng-Chuan Lung)

指導教授 ： 謝淑貞

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2021年)

https://doi.org/10.6342/NTU202100211

摘要

根據衛福部國民健康署的統計，台灣大腸直腸癌的發生率連續十三年位居癌症第一名。大腸直腸癌的發展由良性瘜肉開始，再經過基因突變、危險因子等使之轉變為癌化的腺瘤，最後演變成具有侵略性的腺癌並進行遠端轉移。初期大腸直腸癌五年存活率高達90%，而若發展為遠端轉移，則五年存活率只剩14%。中國橄欖(Canarium album L.)富含多種植化素如gallic acid, ellagic acid, methyl gallate, ethyl gallate等，其果實與其他部位分離出來的純物質已被證實具有許多生理機能效果，例如調節脂質代謝、改善代謝異常、抗氧化等功效，而中國橄欖萃取物對癌細胞的影響也分別在動物與細胞實驗中證實。本實驗室先前研究指出中國橄欖水萃物殘渣甲醇萃取乙酸乙酯區分層(CO-EtOAc, COE) 在異位大腸癌小鼠模式中具有抑制大腸癌細胞增生之功效。細胞模式也確認COE對HCT116人類大腸癌細胞有抑制增生、誘導細胞凋亡的效果，並透過蛋白質體學探討分子機制發現COE會活化p53蛋白以促進細胞週期停滯，進而誘導細胞凋亡。本研究使用LC-MS/MS分析COE內純物質成分以探討COE抑制腸癌細胞增生的功效性成分，發現主要的功效性成分為gallic acid，同時發現其需要COE內其他微量物質以達到近似高劑量COE (400 μg/mL)的效果，另外亦驗證gallic acid加上低劑量COE (25 μg/mL)可藉由誘導p53蛋白以促進細胞凋亡。在轉移部份研究發現COE具有抑制癌細胞遷移(migration)及侵入(invasion)的效果，且具有劑量效應，進一步探討分子機制顯示COE可以抑制Src及FAK蛋白表現量及β-catenin易位進核的現象，且會促進細胞貼盤的能力，可能為COE抑制細胞遷移及侵入的原因。

關鍵字

大腸直腸癌 ； 轉移 ； 中國橄欖萃取物 ； 功效性成分 ； 機制

並列摘要

According to the National Health Administration of the Ministry of Health and Welfare, the incidence of colorectal cancer in Taiwan has ranked first among cancers for 13 years. The development of colorectal cancer begins with benign polyps, and then turns into cancerous adenomas through genetic mutations and risk factors, and finally develops into aggressive adenocarcinomas with distant metastasis. The five-year survival rate for early colorectal cancer is as high as 90%. However, if it develops to distant metastasis, the five-year survival rate is only 14%. Chinese olive (Canarium album L.) is rich in many plant chemicals such as gallic acid, ellagic acid, methyl gallate, ethyl gallate, etc. The pure compounds separated from the fruit and other parts of Chinese olive have many physiological functions, such as regulating lipids metabolism, improving of abnormal metabolism, anti-oxidation, and other functions. The effects of Chinese olive extract on cancer have also been confirmed in animal and cell models. Our laboratory's previous research indicated that the ethyl acetate soluble fraction of methanol extraction from Chinese olive water-insoluble residue showed the inhibitory effect on the proliferation of colorectal cancer cells in a syngeneic mouse model. The cell model results also confirmed that COE inhibited cell proliferation and induced apoptosis in human HCT116 colorectal cancer cells. Through proteomics to explore the molecular mechanism, we claimed that COE can activate p53 protein to promote cell cycle arrest and induce cell apoptosis. In this study, LC-MS/MS was used to analyze the components in COE to explore the functional ingredients of COE for inhibiting the proliferation of human colorectal cancer cells. The main functional ingredient was gallic acid, but it required other tiny compounds in COE to achieve an approximate effect of the high-dose COE (400 μg/mL), and we also verified that gallic acid pulsed low-dose COE (25 μg/mL) could induce p53 protein to promote cell apoptosis. In addition, COE could inhibit the migration and invasion of cancer cells with dose-dependent effect. Further investigation of the molecular mechanism shows that COE could inhibit the expression of Src and FAK protein and translocation of β-catenin into the nucleus, and promote the ability of cells adhesion, which may cause the inhibition of cell migration and invasion of COE.

並列關鍵字

Colorectal cancer ； metastasis ； Chinese olive extract ； functional ingredients ； mechanism

參考文獻

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