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系統識別號 U0007-2607201710460300
論文名稱(中文) 白藜蘆醇抗口腔癌之角色:探討細胞自噬、凋亡與轉移作用
論文名稱(英文) The role of resveratrol in oral cancer: studies on autophage, apoptosis and metastasis
校院名稱 臺北醫學大學
系所名稱(中) 藥學系(碩博士班)
系所名稱(英) School of Pharmacy
學年度 105
學期 2
出版年 106
研究生(中文) 張朝翔
研究生(英文) Chao-Hsiang Chang
電子信箱 vin066@ms14.hinet.net
學號 D301098007
學位類別 博士
語文別 中文
口試日期 2017-07-15
論文頁數 108頁
口試委員 指導教授-王靜瓊
共同指導教授-許光陽
委員-黃聰龍
委員-郭盛助
委員-陳彥州
中文關鍵字 口腔癌  耐藥性  細胞自噬  細胞凋亡  轉移  白藜蘆醇 
英文關鍵字 oral cancer  resistance  autophagy  apoptosis  metastasis  resveratrol 
學科別分類
中文摘要 白藜蘆醇是一種多酚類的天然物具有多樣性藥理活性。目前沒有文獻發表研究白藜蘆醇如何影響順鉑耐藥性口腔癌細胞(CAR)的作用與抑制其侵襲與移轉。結果顯示,相對於正常口腔細胞,白藜蘆醇對CAR具選擇性毒殺作用。白藜蘆醇可造成CAR細胞自噬性死亡與凋亡並透過螢光試劑AO與MDC染色證實其生成酸性囊泡與自噬空泡。同時DNA斷裂與濃縮片段,也可在白藜蘆醇誘發CAR凋亡細胞中觀察到。3-MA (PI3K抑制劑)與Compound c (AMPK抑制劑),抑制白藜蘆醇所誘發CAR細胞自噬囊泡形成、LC-II蛋白量與自噬作用。Z-VAD-FMK (泛caspase抑制劑),抑制白藜蘆醇所誘發CAR細胞之 cleaved caspase-3、cleaved caspase-9含量和與細胞凋亡作用。白藜蘆醇增加CAR細胞AMP kinase磷酸化與調控細胞自噬與前自凋性相關訊息傳遞。此外,白藜蘆醇會刺激CAR細胞自凋性 mRNA基因表現包括 Atg5、Atg12、Beclin-1 和 LC3-II。實驗結果顯示白藜蘆醇亦會抑制CAR細胞侵襲與移行作用。實驗中,白藜蘆醇小於75 µ下不具細胞毒性。白藜蘆醇可透過明膠基質金屬蛋白酶實驗證實降低MMP-2與MMP-9活性與抑制 MPPK訊息包括ERK、JNK與p-38的活化達到抑制CAR細胞侵襲與移行作用。綜合來說,本論文結果顯示白藜蘆醇對耐藥性口腔癌細胞誘發細胞自噬性死亡與凋亡。白藜蘆醇具有對耐藥性口腔癌抑制轉移的抗癌活性。此結果可以供白藜蘆醇未來作為新的營養醫學品協助口腔癌治療。
英文摘要 REV (REV) is one of the polyphenolic components in plants with various pharmacological activities. No report showed how REV affected cisplatin-resistant human oral cancer cell, CAR, and suppression of cell invasion and migration in vitro. Our results demonstrated that REV was low cytotoxic in normal oral cell but not in CAR. It provoked autophagic cell death to form acidic vesicular organelles (AVOs) and autophagic vacuoles by acridine orange (AO) and monodansylc adaverine (MDC) staining. Either DNA fragmentation or DNA condensation occurred in REV-triggered CAR cell apoptosis. These inhibitors of PI3K class III (3-MA) and AMP-activated protein kinase (AMPK) (compound c) suppressed the autophagic vesicle formation, LC3-II protein levels and autophagy induced by REV. The pan-caspase inhibitor Z-VAD- FMK attenuated cleaved caspase-9, cleaved caspase-3 and cell apoptosis. REV also enhanced phosphorylation of AMPK and regulated autophagy- and pro-apoptosis-related signals in CAR cell. Importantly, REV also stimulated the autophagic mRNA gene expression, including Atg5, Atg12, Beclin-1 and LC3-II in CAR cell. Our results demonstrated that REV inhibited cell migration ability by wound healing assay and suppressed CAR cell invasion using the transwell- matrigel assay. Important, REV at less than 75 μM had no cytotoxic effect in CAR cell by MTT assay. REV also attenuated matrix metalloproteinase-2/-9 enzyme activities in CAR cell by gelatin zymography. The suppressive effects of MAPK signaling (ERK, JNK and p38) were observed in REV-treated CAR cell. Overall, our findings indicate that REV is likely to induce autophagic and apoptotic death in drug-resistant oral cancer cell. REV also had suppression of metastatic effect on drug-resistant oral cancer and could be a new nutraceutical to help oral cancer treatment in the near future.
論文目次 誌謝
目錄
圖目錄
表目錄
中文摘要
Abstract
縮寫表
第一章、 緒論
第一節、 口腔癌概述、治療與耐藥性……………………..……1
第二節、 癌細胞之自噬、凋亡與轉移作用……………….18
第三節、 白藜蘆醇相關研究與應用……………………...…22
第四節、 研究目的……………………………………………35
第二章、 實驗材料與方法
第一節、 實驗材料…………………….……………...………...36
第二節、 實驗方法………………………….…………..............43
第三章、 實驗結果
第一節、 白藜蘆醇對CAR誘發之自噬與凋亡作用………….48
第二節、 白藜蘆醇抑制CAR侵襲與移行作用………………..58
第四章、 討論…………………………………………………..…………....64
第五章、 結論與展望………………………………………..…..…………..69
第六章、 參考文獻………………………………………….….....................71
附錄
一、 已發表之SCI論文
二、 擬發表之SCI論文Abstract
三、 CAL 27與CAR之鑑定與比較資料
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