樟芝及其單一萃取物於非小細胞肺癌抗腫瘤之研究

Study on the anti-cancer effect of Antrodia cinnamomea and its single compound on non-small cell lung cancer

陳柏宇(Po-Yu Chen)

指導教授 ： 鄧文炳

臺北醫學大學/口腔醫學院/生醫材料暨工程研究所/碩士(2011年)

https://doi.org/10.6831/TMU.2011.00006

摘要

Antrodia cinnamomea稱為樟芝，為台灣特有藥物真菌，過去的研究中，證實具抗腫瘤生長、抗發炎等藥物活性。本研究利用樟芝酒精萃取物ACAE及其單一萃取物ST-1於人類非小細胞肺癌細胞H441GL轉移機制之探討。實驗設計分為兩大部分，第一部分，於先前本實驗室發現，樟芝酒精萃取物ACAE對非小細胞肺癌生長有良好的抑制增生能力。藉此，本研究更進一步以ACAE於H441GL細胞轉移機制之探討，於基因及蛋白質表現證實ACAE能使H441GL細胞週期停滯於G1時期；於細胞凋亡實驗證實能降低Bcl-2之表現，增加caspase 9 與caspase 3之活性，使DNA產生片斷化，誘導細胞走向細胞凋亡；wound scratch、transwell實驗及降低CXCR4及MMP-2之表現，證實能抑制細胞移動及侵襲行為。第二部分，為了探求樟芝單一抗癌活性成分，進一步分離出樟芝固醇類之單一萃取物ST-1，將肺癌細胞處理48小時，發現在某些濃度下由DNA片斷化實驗證實ST-1可促進肺癌細胞凋亡；wound scratch及transwell實驗證實可抑制細胞移動行為；zymography則發現有抑制細胞侵襲活性；基因層級上，證實可調控細胞轉移相關基因E-cadherin、vimentin、CXCR4與MMP-2及細胞週期相關基因cyclin A與cyclin E之表現。由該結果證實，ST-1可藉由透過抑制細胞移動、侵襲及促其細胞凋亡而達到抑制肺癌細胞H441GL轉移的能力。

關鍵字

樟芝酒精萃取物；非小細胞肺癌；細胞轉移；細胞週期；ST-1

並列摘要

In this study, we evaluated the anti-metastatic effects of Antrodia cinnamomea alcohol extracts (ACAE) and its single compound ST-1 on non-small cell lung cancer (H441GL cells). The study consists of two main parts. In the first part, consistent with our lab’s previous discovery, ACAE inhibited the growth of non-small cell lung cancer. This study further focus on anti-metastatic effects. Through gene and protein analysis, ACAE induced cell cycle arrest at G1 phase. At cell apoptotic analysis, we demonstrated that it induced cell apoptosis by down-regulate Bcl-2 expression and increased the activity of caspase 9 and caspaase 3. Through wound scratch, transwell assay and MMP-2 activity, we demonstrated that it inhibited the cell migration and invasion ability. The second part, we further extracted ST-1 from Antrodia cinnamomea, the steroid compound and treated lung cancer cells for 48 hours. Through DNA fragmentation, we demonstrated that ST-1 led to H441GL cell apoptosis. Through both wound scratch and transwell assays, we demonstrated that it limited cell migration. Through zymography, we demonstrated that it inhibited cell invasion activity. At the genetic level, we demonstrated that it controlled the E-cadherin, vimentin, CXCR4, and MMP-2 genes associated with cell metastasis and the cyclin E and cyclin A genes associated with the cell cycle. These experiments demonstrated that ST-1 limited the metastasis of H441GL by limiting H441GL migration and invasion activity and by encouraging H441GL apoptosis.

並列關鍵字

Antrodia cinnamomea alcohol extract (ACAE)；non-small cell lung cancer；metastasis；cell cycle；ST-1

參考文獻

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