根據美國癌症協會估計，肝癌在男性癌症死亡排名中位居第五，在女性癌症死亡排名中位居第八。而根據台灣衛福部統計，肝癌在台灣的癌症死亡排名中甚至連續幾年都排行第二名。肝癌的死亡率如此之高是由於肝臟神經在內部少有，所以癌症產生時不易察覺，察覺時也都到了中晚期，且肝癌的五年存活率非常低，增加了治療上的難度。因此在本研究中的主旨是希望能發現抑制肝癌的方法，並用蛋白質體學的方式來探討其抑制的機制。
牛樟芝 (Antrodia Cinnamomea)是一種真菌，原只生長於台灣的牛樟樹木中，在民間被當作解肝毒的一種食材，在學術研究上也被證實其成分之一的萜類化合物 (terpenoids)具有抑制肝癌細胞增生的能力。所以在實驗中首先以乙醇提取牛樟芝 (ethanolic extracts of Antrodia Cinnamomea, EEAC)，接著分別加入正常肝細胞株Chang Liver和肝癌細胞株C3A、HepG2中進行細胞存活率試驗 (MTT assay)。然後用蛋白質體學的方式去分析EEAC的效用，包括二維差異電泳 (2D-DIGE)、差異分析軟體DeCyder、介質輔助雷射脫附游離/飛行時間質譜 (MALDI-TOF-MS)找出差異之蛋白質。
透過SwissProt資料庫和KEGG路徑分析，將這些鑑定出來的蛋白質依據其功能與表現量進行分類，觀察到許多蛋白質在蛋白質摺疊、氧化還原調節、醣解作用的功能中表現量下降，因此我們推測EEAC會導致癌細胞產生內質網壓力 (ER stress)，進而誘導細胞凋亡。為了證實這個推測，我們進行了西方點墨法 (western blot)去壓跟內質網壓力相關的抗體，結果證實EEAC是透過蛋白質摺疊錯誤與細胞氧化還原不平衡所引起的內質網壓力來抑制癌細胞，導致細胞產生毒性而凋亡。

Of all the cancers discovered in patients living in Taiwan, liver cancer is the prevalent type of cancer diagnosed in patients. The American Cancer Society estimates liver cancer is the 5th leading cause of death in males and the 8th in females in the United States. Thus, our objective is to discover effective treatment options for liver cancer.
In this study, three cell lines are used, one normal liver cell lines, Chang Liver, and two hepatocellular carcinoma cell lines, C3A and HepG2. First, we perform MTT assays with ethanolic extracts of Antrodia Cinnamomea (EEAC), a Taiwan endemic fungus species used as an Asian folk medicine. Then, we use proteomic analysis including two-dimensional differential gel electrophoresis (2D-DIGE), DeCyder differential analysis software, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and immunoblotting to verify differentially expressed proteins. By searching SwissProt and KEGG pathway analysis, these identified proteins can be classified according to their subcellular locations, functional ontology, and expression profiles. In view of observing numerous proteins with their biological functions in protein folding, we performed immunoblotting to stain ER related antibodies specific to the target protein.
To sum up, our results indicated EEAC promote cytotoxicity in liver cancer cells rather than normal liver cells through ER-stress caused by protein misfolding and cellular redox unbalance.

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