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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳振陽(Jen-Yang Chen) | |
| dc.contributor.author | Yu-Chia Chuang | en |
| dc.contributor.author | 莊育嘉 | zh_TW |
| dc.date.accessioned | 2021-06-13T07:50:22Z | - |
| dc.date.available | 2008-08-02 | |
| dc.date.copyright | 2005-08-02 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-25 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36050 | - |
| dc.description.abstract | 根據血清學與細胞遺傳學等多項證據，顯示EB病毒(Epstein-Barr virus)與鼻咽癌的形成有著密切的關係。而在EB病毒相關的腫瘤組織中，往往可以發現異常的染色體型態，因此，EB病毒的感染是否會造成基因體的不穩定，最終導致細胞的癌化，便成為值得探討的問題。基因體不穩定所引發的細胞轉形，其原因有許多：如DNA修復作用的失常、細胞週期檢查系統遭受干擾，細胞凋亡的抑制等。目前已知，某些病毒感染後，會影響到細胞DNA修復系統，促使細胞轉形。本實驗室先前已證實了，在宿主再活化試驗中，EB病毒第一型潛伏膜蛋白 (latent membrane protein 1, LMP1) 能夠抑制上皮細胞的DNA修復作用；藉由即時定量反轉錄PCR的分析結果，亦發現LMP1會抑制某些DNA修復基因的表現，包括Cockayne syndrome group B (CSB)和Xeroderma pigmentosum complementation group A (XPA)。CSB與XPA參與著核苷酸切除修復機制 (nucleotide excision repair, NER)，能夠移除因UV照射而毀損的DNA，避免細胞發生轉錄上的障礙。在NER中，XPA主要負責辨識受到傷害的DNA，CSB則與轉錄伴隨修復機制 (transcription-coupled repair) 有關，負責吸引其他的NER蛋白。
本研究欲探討LMP1能否調控CSB和XPA的表現，希望藉此解釋LMP1影響DNA修復機制的途徑。在mRNA層面，不論是短暫轉染LMP1表現質體至H1299/bcl2細胞中，或是誘發LMP1在RHEK/tet-LMP1細胞的表現，皆能證實LMP1的抑制效果。在蛋白質層面，本研究則偵測到當細胞內有LMP1的存在，其CSB和XPA的蛋白質表現也會有減少的傾向。 為了瞭解LMP1的抑制現象是否藉由影響CSB和XPA啟動子活性來達成，經由PCR的方法，我們構築了一系列CSB和XPA刪除變異啟動子於pGL3-basic冷光酵素報告質體。透過冷光酵素分析，發現在數種上皮細胞中，隨著LMP1量的提高，CSB和XPA啟動子的活性隨之下降，但卻無法界定出LMP1的反應區域。此結果顯示LMP1的反應區域可能位在靠近轉錄起始的位置。 為了進一步找出LMP1的轉抑制區，使用一系列的LMP1突變株進行冷光酵素分析。結果顯示LMP1主要以CTAR1 (C-terminus activating region 1) 抑制CSB和XPA啟動子，且以CTAR2 (C-terminus activating region 2) 協同抑制XPA啟動子活性。CTAR1和CTAR2負責NF-B、MAPK/p38和PI3K/Akt訊息傳遞路徑，但LMP1是否透過這些訊息傳遞路徑而影響DNA修復基因的啟動子，則有待進一步的研究來證實。 本研究證實LMP1抑制CSB和XPA啟動子的活性，藉以減少細胞內CSB和XPA mRNA的含量，並抑制CSB和XPA的蛋白表現，但LMP1是否藉此而影響DNA修復作用，則展望後續的實驗來闡明。 | zh_TW |
| dc.description.abstract | Serological and cytogenetic studies show that Epstein-Barr virus (EBV) is closely related to nasopharyngeal carcinoma. Many chromosomal abnormalities were observed in the tissues of EBV-associated malignancies. It implies that EBV infection may induce genomic instability which is highly associated with the defects of DNA repair, cell cycle checkpoint and apoptosis. Latent membrane protein 1 (LMP1), an oncoprotein encoded by EBV, is an integral membrane protein, which acts like a constitutively active tumor necrosis factor receptor. LMP1 is essential for EBV-mediated B cell transformation and oncogenic to rodent fibroblasts. It regulates transcription factor activity and gene expression through signaling pathways ,including NF-B, MAPK/p38, and PI3K/Akt signaling pathways, and therefore further investigation is needed to identify which pathway plays the important role.
In sum, our study showed that LMP1 suppresses mRNA expression of CSB and XPA by inhibiting promoter activity and represses protein expression of CSB and XPA. However, to understand whether LMP1 represses DNA repair by reducing the expression of both CSB and XPA or other DNA repair gene needs further study. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T07:50:22Z (GMT). No. of bitstreams: 1 ntu-94-R92445128-1.pdf: 1823056 bytes, checksum: ea42deafa220f1db7f8c1e15748b6291 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 中文摘要 1
英文摘要 2 1. 序論 3 2. 實驗目的 16 3. 材料與方法 17 4. 實驗結果 27 5. 討論 32 6. 參考文獻 37 7. 圖表 49 8. 附錄 61 | |
| dc.language.iso | zh-TW | |
| dc.subject | DNA修復作用 | zh_TW |
| dc.subject | EB病毒 | zh_TW |
| dc.subject | 第一型潛伏膜蛋白 | zh_TW |
| dc.subject | 啟動子 | zh_TW |
| dc.subject | promoter | en |
| dc.subject | DNA repair | en |
| dc.subject | EBV | en |
| dc.subject | LMP1 | en |
| dc.subject | CSB | en |
| dc.subject | XPA | en |
| dc.title | EB病毒第一型潛伏膜蛋白對CSB和XPA啟動子活性的影響 | zh_TW |
| dc.title | The effect of Epstein-Barr Virus latent membrane protein 1 on CSB and XPA promoter activity | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王萬波,董馨蓮,李財坤 | |
| dc.subject.keyword | EB病毒,第一型潛伏膜蛋白,啟動子,DNA修復作用, | zh_TW |
| dc.subject.keyword | EBV,LMP1,CSB,XPA,promoter,DNA repair, | en |
| dc.relation.page | 64 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-26 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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