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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 郭彥彬 | |
dc.contributor.author | Chia-Yun Huang | en |
dc.contributor.author | 黃佳云 | zh_TW |
dc.date.accessioned | 2021-06-17T02:11:02Z | - |
dc.date.available | 2020-02-22 | |
dc.date.copyright | 2018-02-22 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-01-19 | |
dc.identifier.citation | Aitken, R.J., and Roman, S.D. (2008). Antioxidant systems and oxidative stress in the testes. Oxid Med Cell Longev 1, 15-24.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/67997 | - |
dc.description.abstract | 衛生福利部今年公佈的 103 年癌症登記報告指出,口腔癌為臺灣十大癌症發生率的第五位,也是台灣年增率最高癌症。臺灣口腔癌發生與嚼檳榔有密切關係,但檳榔致癌機制仍未詳細闡明。文獻指出檳榔中的主要成分檳榔鹼(arecoline)是主要導致口腔癌發生的致病因子。八聚體結合轉錄因子4(Oct4)是口腔癌中幹細胞基因,研究發現OCT4的表現與口腔癌病人的預後效果呈高度負相關性,但Arecoline是否可誘導口腔上皮細胞OCT4表現及其機轉並不清楚。在本篇研究,我們發現Arecoline會誘導牙齦正常上皮SG細胞、口腔癌CA9-22及SAS細胞株中OCT4的表現。競爭性乙醯膽鹼蕈毒接受器(muscarinic acetylcholine receptor)抑制劑atropine及M4 muscarinic acetylcholine receptor拮抗劑Tropicamide可降低由Arecoline所誘導的OCT4的表現。TGF-β1中和抗體、ALK5抑制劑(SB431542)以及Smad3抑制劑(SIS3)可降低Arecoline誘導SG及Ca9-22細胞中OCT4的表現,顯示Arecoline 經由TGF-β1訊息傳遞路徑誘導口腔上皮細胞OCT4的表現。進一步發現Arecoline可以增加SG及Ca9-22細胞培養液中活化態TGF-β1的表現量。前處理抗氧化劑NAC能夠明顯降低Arecoline誘導SG及Ca9-22細胞的活化態TGF-β1。抗氧化劑NAC、PEG-catalase、mitochondria-targeted O2-抑制劑(Mito-TEMPO)、NO抑制劑(L-NAME)以及MnTBAP能夠明顯抑制Arecoline誘導SG及Ca9-22細胞活化態TGF-β1的表現量。EGCG可抑制Arecoline誘導SG及Ca9-22細胞活化態TGF-β1的表現量。 EGFR抑制劑(AG1478)、PI3K抑制劑(LY294002)、P38/ MAPK抑制劑(SB203580)、JNK抑制劑(SP600125)能夠明顯降低TGF-β1誘導SG及Ca9-22細胞中OCT4的表現。此外我們發現兒茶素(EGCG)對於檳榔鹼誘導誘導SG及CA9-22細胞中OCT4的表現皆有明顯的抑制效果。這些結果顯示,Arecoline經由Muscarinic receptor、ROS以及TGF-β1路徑誘導OCT4的表現,並得知此誘導路徑可藉由EGCG抑制Arecoline誘導細胞產生OCT4。 | zh_TW |
dc.description.abstract | In Taiwan, oral cancer is the forth leading cause of death in males 2011. Areca nut (AN) chewing is the most important risk factor of oral cancer. Octamer-binding transcription factor 4 (OCT4) has been shown to play a central role in the pathogenesis of many human cancers. Chiou et al. reported that patients with higher OCT4 expression have worse overall survival. We observed Arecoline, a main alkaloid found in AN, increased OCT4 synthesis in a dose- and time- dependent manner in SG, Ca9-22 and SAS cell. Pretreatment with muscarinic receptor inhibitor (Atropine and Tropicamide) significantly reduced Arecoline induced OCT4 synthesis. We aimed to examine the possible signal transduction pathways involved in TGF-β1-induced OCT4 expression in SG and Ca9-22 cell, we found the effect that Arecoline induces OCT4 activation was inhibited by TGF-β1 neutralizing antibody, ALK5 inhibitor(SB431542) and Smad3 inhibitor(SIS3). We have also shown that EGFR inhibitor(AG1478), PI3K inhibitor(LY294002), P38/MAPK inhibitor(SB203580), and JNK inhibitor(SP600125) significantly abrogate the TGF-β1-induced levels of OCT4 protein, indicating ALK5/Smad3, EGFR, PI3K, P38/MAPK, and JNK are involved in the TGF-β1-induced OCT4 in SG and Ca9-22 cells. Furthermore, epigallocatechin-3-gallate (EGCG) completely inhibited Arecoline-induced OCT4 synthesis and inhibition is dose-dependent. We found Arecoline increased activated TGF-β1 level in SG and Ca9-22 cells. Pretreatment with antioxidant NAC significantly reduced Arecoline-induced activated TGF-β1 level in SG and Ca9-22 cells. We further found antioxidant NAC, PEG-catalase, mitochondria-targeted O2- inhibitor(Mito-TEMPO), NO inhibitor(L-NAME) and MnTBAP significantly inhibited Arecoline-induced activated TGF-β1 level in SG and Ca9-22 cells. In addition, EGCG significantly inhibited Arecoline-induced activated TGF-β1 level in SG and Ca9-22 cells. Taken together, our results indicate the Arecoline enhances cancer stem cell marker by increasing OCT4 expression via muscarinic receptor, ROS and TGF-β1 pathways, and EGCG potentially qualifies as a useful reagent for the prevention and therapy of oral cancer. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T02:11:02Z (GMT). No. of bitstreams: 1 ntu-107-R04450003-1.pdf: 2204366 bytes, checksum: ce2825937cfde1c0e466a5d3b10da260 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 口試委員會審定書……….………………………………………………………….Ⅰ
謝誌………………………………….……………………………………………….Ⅱ 中文摘要…………………………….………………………………………….……Ⅲ Abstract………………………………………………………………………………Ⅴ 第一章 導論…………………………………………………………………………..1 第一節 口腔癌…………………………………………………………...…….1 1-1 口腔癌的簡介…………………………………………………...…..…1 1-2 口腔癌的診斷與分期………………………………………...………..2 1-3 口腔癌治療………………………………………………...…………..3 第二節 檳榔鹼(Arecoline)……………………………………………………6 2-1 Arecoline的訊息傳遞路徑……………………...………...…….……...6 2-2 Arecoline與口腔癌…………………………….…………………....….7 第三節 轉化生長因子(TGF-β1)……………………………………………9 3-1 TGF-β1的簡介…………………………………………….…..……….9 3-2 TGF-β1的訊息傳遞路徑…………………………………………….10 3-3 TGF-β1與癌症………………………………………………………..12 第四節 八聚體結合轉錄因子4 (OCT4)………………………………….14 4-1 OCT4的簡介………………………………………………………….14 4-2 OCT4與癌症………………………………………………………….15 第五節 活性氧化物(ROS)…………………………………………………..16 5-1 ROS的簡介…………………………………………………………...16 5-2 ROS與癌症…………………………………………………………...17 第六節 兒茶素(EGCG)……………………………………………………...19 第二章 實驗目的…………………………………………………………..………21 第三章 材料與方法………………………………………………………………..22 第一節 細胞株與細胞培養…………………………………………………..22 1-1 細胞株的培養………………………………………………………...22 1-2 繼代培養……………………………………………………………...22 第二節 藥物處理……………………………………………………………..23 2-1 種細胞以及starvation………………………………………………..23 2-2 抑制劑、抗氧化劑以及中和抗體使用資料………………………….23 第三節 西方點墨法…………………………………………………………..26 3-1 蛋白質萃取…………………………………………………………...26 3-2 配置膠體……………………………………………………………...26 3-3 電泳分析……………………………………………………………...26 3-4 蛋白質的轉漬……………………………………………………...…27 3-5 抗體的使用…………………………………………………………...27 3-6 顯影呈色………………………………………………………….......28 第四節 Enzyme-linked immunosorbent assay(ELISA)……………………..29 4-1 Coating………………………………………………………………...29 4-2 Blocking…………………………………...…………………………..29 4-3 Sample and standard…………………………………………………..29 4-4 Detection………………………………………………………………29 4-5 Streptavidin-HRP……………………………………………………...30 4-6 TMB substrate…………………………………………………………30 4-7 Stop reaction & read OD value………………………………………..30 第五節 統計分析…………………………………………………………..…31 第四章 結果………………………………………………………………………..32 Arecoline誘導Ca9-22、SG及SAS細胞OCT4蛋白質表現…………………..32 Arecoline經由MR路徑誘導Ca9-22及SG細胞OCT4蛋白質表現………….32 Arecoline經由TGF-β路徑誘導Ca9-22及SG細胞OCT4蛋白質表現………33 TGF-β1誘導Ca9-22及SG細胞OCT4蛋白質表現…………………………...33 EGFR、PI3K、P38/MAPK、JNK調控TGF-β1誘導Ca9-22及SG細胞OCT4表現……………………………………………………………………………..33 EGCG抑制Arecoline誘導Ca9-22、SG及SAS細胞OCT4蛋白質表現……...34 Arecoline誘導Ca9-22及SG細胞細胞釋出活化態TGF-β1但不涉及TGF-β1生和成…………………………………………………………………………..34 MMP不涉及Arecoline誘導Ca9-22及SG細胞細胞釋出活化態TGF-β1…...35 Arecoline經由MR誘導Ca9-22及SG細胞細胞釋出活化態TGF-β1………..35 不同ROS抑制劑對於Arecoline誘導Ca9-22及SG細胞細胞釋出活化態TGF-β1的抑制能力…………………………………………………………….35 EGCG抑制Arecoline誘導Ca9-22及SG細胞細胞釋出活化態TGF-β1…….36 第五章 討論………………………………………………………………………..37 第六章 圖與表……………………………………………………………………..41 第七章 References…………………………………………………………………52 | |
dc.language.iso | zh-TW | |
dc.title | Arecoline誘導口腔上皮細胞OCT4表現及其機轉 | zh_TW |
dc.title | Arecoline induces OCT4 exprseeion in oral epithelial cells and its mechanism | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張瑞青,鄭世榮 | |
dc.subject.keyword | 檳榔鹼,口腔癌,八具體結合轉錄因子4,活性氧化物,兒茶素,轉化生長因子, | zh_TW |
dc.subject.keyword | Arecoline,oral cancer,octamer-binding transcription factor 4,Reactive oxygen species,Epigallocatechin-3-gallate,Transforming growth factor-β1, | en |
dc.relation.page | 66 | |
dc.identifier.doi | 10.6342/NTU201800111 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-01-22 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 口腔生物科學研究所 | zh_TW |
顯示於系所單位: | 口腔生物科學研究所 |
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