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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳信銘(Hsin-Ming Chen) | |
| dc.contributor.author | Chia-Yin Chin | en |
| dc.contributor.author | 秦嘉霠 | zh_TW |
| dc.date.accessioned | 2021-06-07T17:57:03Z | - |
| dc.date.copyright | 2012-09-19 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-08-14 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/15979 | - |
| dc.description.abstract | 口腔黏膜下纖維化症(Oral submucous fibrosis，OSF)是一種持續性發炎所導致的口腔黏膜病變，屬於潛在性癌前障礙，主要因口腔黏膜下組織層過多的膠原蛋白堆積，而造成纖維化。已知在發炎的過程中，部分發炎前細胞激素會被大量分泌，其中像是IFN-γ、TGF-β等，此些激素參與纖維化的致病機轉已被進一步研究，另外，在之前的文獻中指出，B7-H1蛋白表現會受到TGF-β、IFN-γ、TNF-α等細胞激素所調控，因此，在本研究中，其主要目的是在探討B7-H1於口腔黏膜下纖維化症之致病機轉中，其所扮演的角色。
首先，利用B7-H1抗體進行免疫組織化學染色，偵測在正常口腔黏膜組織(normal oral mucosa；NOM)及口腔黏膜下纖維化症組織(oral submucosa fibroblast；OSF)的切片中B7-H1表現的情形，結果發現，在OSF的切片中明顯看到部分表現B7-H1的纖維母細胞靠近上皮層組織，且同時觀察到在上皮組織層中B7-H1的表現也較正常組織中多。其次，正常黏膜纖維母細胞以arecoline處理後，細胞中B7-H1表現呈現增加的趨勢，並且發現用JNK抑制劑可抑制B7-H1的表現。過去文獻指出在人類皮膚癌中，B7-H1的過度表現會去調控上皮-間質轉換現象(EMT)，而本研究免疫組織化學染色結果也發現，在OSF組織中，表現B7-H1的纖維母細胞大多集中在上皮下結締組織，推測可能有EMT現象產生而導致纖維母細胞的增生活化。利用細胞實驗看in vitro中B7-H1表現，用TGF-β去處理S-G上皮細胞，發現在S-G細胞中EMT 生物標記蛋白增加且B7-H1表現也增加，處理JNK抑制劑可以抑制B7-H1的表現，並且發現EMT biomarker蛋白也有減少的趨勢，進一步用B7-H1 shRNA來確認B7-H1是否會進一步去調控EMT，結果顯示當抑制B7-H1表現時，會影響EMT相關蛋白的表現。 | zh_TW |
| dc.description.abstract | Oral submucous fibrosis is a continuous, chronic, insidious and inflammatory disease of oral mucous that is a kind of potentially malignant disorders. OSF is mainly due to consumed areca quid which is lead to fibrosis in the oral cavity.
In the progression of inflammatory, some pro-inflammatory and pro-fibrotic cytokine could be upregulated, like TGF-β、IFN-γ, etc., the exact pathogenesis of OSF and cytokine was still unclear and needed to study .However, the previously study had showed that B7-H1 was regulated by IFN-γ in the human dermal fibroblast. Therefore, in this study, we investigated whether B7-H1 was involved in the mechanisms of OSF diseases. First, we investigated the expression of B7-H1 in normal oral mucosa tissue and in OSF patient’s tissue by immunohistochemistry (IHC).The result showed that most of fibroblasts that express B7-H1 was close to epithelium layer in OSF tissue. It also showed that the expression of B7-H1 in epithelium layer in OSF is much more than in NOM. Then, we treated NOM fibroblast with arecoline and analyzed the expression of B7-H1 by Western blot. The result showed that B7-H1 is up-regulated by arecoline in NOM, and JNK inhibitor could reduce B7-H1 expression. The reference indicated that B7-H1 over-expression could be regulated Epithelial-mesenchymal transition (EMT) in human skin cancer. The result of IHC showed that fibroblasts that express B7-H1 were in the juxta-epithelial connective tissue. The reference showed that EMT is one of important mechanisms of OSF, and to investigate whether B7-H1 is involved in EMT which is lead to fibrosis. We treated S-G epithelial cell with TGF-β, and the result showed that EMT biomarker protein and B7-H1 expression was increased. And then, we pre-treat S-G cell with JNK inhibitor and then treat with TGF-β, the result showed that the expression of B7-H1 and some EMT biomarker both were reduced. And then, we used B7-H1 specific siRNA to knockdown its expression in S-G cells. The result showed that knockdown of expression of B7-H1 influenced EMT biomarker gene expression. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-07T17:57:03Z (GMT). No. of bitstreams: 1 ntu-101-R99450005-1.pdf: 2443374 bytes, checksum: c9a797dec52385ce702b34778175ec7c (MD5) Previous issue date: 2012 | en |
| dc.description.tableofcontents | 中文摘要...........................................................................................................................1
Abstract..............................................................................................................................2 第一章 導論.....................................................................................................................4 第一節 口腔黏膜下纖維化症.................................................................................4 1-1 口腔黏膜下纖維化症的簡介....................................................................4 1-2 口腔黏膜下纖維化症的致病機轉............................................................5 1-3 口腔黏膜下纖維化症的文獻回顧............................................................6 第二節B7-H1(CD274、PD-L1)..............................................................................8 2-1 B7-H1的簡介.............................................................................................8 2-2 B7-H1的文獻回顧.....................................................................................9 第三節 上皮- 間質細胞轉換（Epithelial-mesenchymal transition，EMT）...11 第二章 實驗目的...........................................................................................................15 第三章 實驗材料與方法...............................................................................................16 第一節 實驗組與控制組檢體的收集...................................................................16 第二節 免疫組織化學染色法 ( Immunohistochemical stain ) ...........................16 第三節 人類頰黏膜纖維母細胞之初代培養.......................................................18 第四節 人類頰黏膜纖維母細胞之繼代培養.......................................................19 第五節 Smulow-Glickman gingival cell line (S-G cell)之繼代培養…..............19 第六節 藥物的處理...............................................................................................19 第七節細胞毒性試驗.............................................................................................20 第八節西方墨點法.................................................................................................20 第九節Reverse tanscriptase PCR...........................................................................21 第十節 shRNA 之使用.........................................................................................22 第十一節 統計方法...............................................................................................22 第四章 結果...................................................................................................................23 OSF 組織切片比NOM組織切片表現較多B7-H1............................................23 免疫組織的化學染色統計結果分析.....................................................................23 不同濃度的Arecoline影響NOM、S-G細胞生長的情形.................................23 Arecoline會誘導NOM細胞B7-H1的表現量增加…........................................24 JNK抑制劑會抑制Arecoline誘導B7-H1在NOM細胞內的表現..................25 Arecoline會誘導B7-H1在S-G細胞的表現量增加...........................................25 TGF-β會誘導B7-H1在S-G細胞的表現量增加...............................................25 JNK抑制劑會抑制B7-H1在S-G細胞內的表現...............................................26 在B7-H1轉染的SG 細胞，抑制EMT biomarker表現...................................26 Snail蛋白的免疫組織化學染色............................................................................27 第五章 討論...................................................................................................................29 第六章 總結...................................................................................................................33 第七章 圖與表...............................................................................................................35 第八章 Reference...........................................................................................................52 | |
| dc.language.iso | zh-TW | |
| dc.subject | 上皮-間質轉換現象(EMT) | zh_TW |
| dc.subject | B7-H1 | zh_TW |
| dc.subject | 口腔黏膜下纖維化症(OSF) | zh_TW |
| dc.subject | oral submucous fibrosis(OSF) | en |
| dc.subject | B7-H1 | en |
| dc.subject | EMT | en |
| dc.title | 口腔黏膜下纖維化症中B7-H1表現之研究 | zh_TW |
| dc.title | The Research of B7-H1 Expression in
Oral Submucous Fibrosis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 郭彥彬(Yen-Ping Kuo),江俊斌(Chun-Pin Chiang) | |
| dc.subject.keyword | 口腔黏膜下纖維化症(OSF),B7-H1,上皮-間質轉換現象(EMT), | zh_TW |
| dc.subject.keyword | oral submucous fibrosis(OSF),B7-H1,EMT, | en |
| dc.relation.page | 55 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2012-08-14 | |
| dc.contributor.author-college | 牙醫專業學院 | 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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