||The role of resveratrol in oral cancer: studies on autophage, apoptosis and metastasis
||School of Pharmacy
||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。
||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.
三、 CAL 27與CAR之鑑定與比較資料
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