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DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 葉秀慧 | |
dc.contributor.author | Chen-Yu Li | en |
dc.contributor.author | 李承彧 | zh_TW |
dc.date.accessioned | 2021-05-19T17:50:44Z | - |
dc.date.available | 2022-09-08 | |
dc.date.available | 2021-05-19T17:50:44Z | - |
dc.date.copyright | 2017-09-08 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-08-15 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7704 | - |
dc.description.abstract | B型肝炎病毒(HBV)相關肝癌有顯著男性發生率高於女性的現象。實驗室先前研究針對HBV相關肝癌DNA進行NGS分析,探討是否存在任何男女差異的現象,結果指出HBV嵌入TERT promoter區域與TERT promoter -124, -146位點發生mutation之兩種活化TERT的機制,均有顯著好發於男性病患的現象。有鑑於實驗室先前研究發現雄激素路徑會經由受體直接結合至病毒基因體enhancer I區域,促進HBV轉錄,而雌激素路徑會藉由受體螯合HNF4α抑制HBV轉錄,造成男性病患較高之病毒量,為可能導致HBV相關肝癌男女差異的一種機制。本研究因此提出探討性激素,包括雄激素路徑及雌激素路徑,是否亦參與在調控上述兩種活化TERT的基因體變異,造成TERT於男性較高表現之可能假說。
本實驗提出利用HBV嵌入TERT promoter區域及包含TERT promoter mutation之報導基因,於細胞培養模式驗證上述假說。針對HBV-TERT promoter integration報導基因所進行之實驗結果顯示,雄激素路徑會促進而雌激素路徑則會抑制報導基因之轉錄。進一步發現將調控HBV轉錄之重要轉錄因子HNF4α敲落後,性激素造成的差異變得不明顯,顯示HNF4α為造成性激素路徑於此報導基因轉錄差異之重要因子。此外針對TERT promoter mutation報導基因所進行之實驗結果顯示,雄激素路徑會促進其轉錄活性,而雌激素路徑較無明顯之影響,而上述作用在將轉錄因子GABPα敲落後即顯著降低,顯示性激素活化此報導基因的機制是建立在GABPα之上。我們實驗結果進一步也發現HNF4α對於GABPα促進此報導基因轉錄上扮演重要的角色,且其促進轉錄作用和雄激素路徑可能為兩獨立調控機制。上述實驗結果初步支持性激素參與調控上述兩種活化TERT的基因體變異,造成TERT於男性較高表現之可能。 | zh_TW |
dc.description.abstract | Hepatitis B virus (HBV) induced hepatocellular carcinoma (HCC) occurs predominantly in male patients. Aided by the capture-NGS, our recent study showed that the insertional mutagenesis to active TERT oncogene occurs preferentially in the male HCC. Meanwhile, the TERT promoter mutations at -124G>A and -146G>A, the most frequent somatic mutation identified in HCC, also occurred preferentially in male HCC. We have previously identified androgen and estrogen effects on the transcription of HBV via targeting to viral enhancer I, leading to higher viral titer in male carriers. This study proposed to investigate if sex hormones can also through the HBV integrated in the host genome or frequent TERT promoter mutations to regulate the expression of TERT oncogenes, as a mechanism contributing to male HCC.
We propose to approach this by the cell culture based TERT-promoter-Luc reporter assay, containing the HBV integration or -124G>A or -146G>A mutations at promoter region. We first found a significantly higher Luc activity from HBV/TERT-Luc than that from WT/TERT-Luc construct, which can be further increased by AR but suppressed by ER pathway. Knockdown of HNF4Α abolished the difference, indicating the critical role of HNF4Α in this regulation. Next, we found a significant elevation in both G(-124)A and G(-146)A-Luc constructs relative to the WT-TERT-Luc, which is dependent on the GABPα. The effect of AR and ERα pathway was examined, showing the reporter activity to be further elevated by AR, in a ligand independent manner, which however was not affected by the ERα pathway. Interestingly, or results showed that HNF4Αalso contributed to the activation of G(-124)A-TERT-Luc. The function of AR however is independent of HNF4Α. Our study thus supported the involvement of sex hormones in regulating the transcription of TERT through HBV integration or frequent TERT promoter mutations. | en |
dc.description.provenance | Made available in DSpace on 2021-05-19T17:50:44Z (GMT). No. of bitstreams: 1 ntu-106-R04445125-1.pdf: 2339870 bytes, checksum: c0a8506759621f24b7ac281fa2ef4044 (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 口試委員審定書 I
致謝 II 摘要 III Abstract IV 目錄 V 第一章 序論 1 1. 肝癌 (Hepatocellular carcinoma) 1 1-1. 肝癌致癌危險因子 1 1-2. 從慢性發炎到肝癌之疾病進程及基因體變異 1 1-3. 肝癌之男女性別差異特性 2 2. B型肝炎病毒 (Hepatitis B virus , HBV) 3 2-1. B型肝炎病毒之結構及生活史 3 2-2. HBV相關肝癌之致病機制 5 3. HBV之轉錄調控機制 6 3-1. 細胞中特定轉錄因子調控HBV RNA之生合成 6 3-2. 性激素對HBV RNA之轉錄之影響 6 3-2-1. 雄激素路徑會促進HBV RNA之轉錄 7 3-2-2. 雌激素路徑會抑制HBV RNA之轉錄 7 4. 肝癌中端粒酶 (Telomerase) 之表現及功能 8 4-1. 端粒 (telomere) 與端粒酶 (telomerase) 8 4-2. 端粒酶在癌細胞中之表現及功能 9 4-3. TERT在肝癌中之活化 9 第二章 材料與方法 11 細胞培養 (Cell culture) 11 質體轉型(Transformation) 11 質體製備 12 細胞轉染(DNA transfection) 12 含有sh-RNA之lentivirus病毒顆粒製備 13 病毒感染 14 細胞均質液之製備 14 蛋白質定量 14 SDS-PAGE 蛋白質膠體電泳 15 西方墨點法 15 冷光酶測定分析(Dual-luciferase reporter assay) 16 統計方法 17 第三章 結果 18 探討B型肝炎相關肝癌中HBV-TERT integration及 TERT promoter mutation 之男女差異 18 報導基因實驗使用之質體 19 探討性激素路徑對於HBV-TERT promoter integration之轉錄調控 20 HNF4α為性激素路徑造成HBV-TERT promoter integration轉錄調控之重要因子 22 探討性激素路徑對於TERT promoter mutation之轉錄調控 23 GABPα為雄性激素路徑活化TERT promoter mutation轉錄作用之重要因子 25 HNF4α為肝細胞中促進TERT promoter mutation轉錄作用之重要因子 26 第四章 討論 27 實驗的侷限性 27 性激素對於TERT promoter mutation轉錄作用調控與臨床現象 27 HNF4α於TERT promoter mutation之調控 28 TERT promoter mutation男女差異只在B型而非C型肝炎相關肝癌 29 附圖 30 參考資料 42 | |
dc.language.iso | zh-TW | |
dc.title | 探討肝癌中性激素路徑對於TERT基因表現之調控 | zh_TW |
dc.title | The effect of sex hormones on TERT expression in hepatcarcinogenesis | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳培哲,鄧述諄 | |
dc.subject.keyword | 性激素,TERT啟動子突變,HBV嵌入,肝癌, | zh_TW |
dc.subject.keyword | Sex hormone,TERT promoter mutation,HBV integration,Hepatocellular carcinoma., | en |
dc.relation.page | 47 | |
dc.identifier.doi | 10.6342/NTU201703348 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2017-08-15 | |
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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