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研究生:沈家興
研究生(外文):Chia Hsing Shen
論文名稱:甲狀腺素受體負向調控lnog non-coding RNA SNHG1進而抑制肝癌細胞的生長
論文名稱(外文):Long non-coding RNA SNHG1 regulated by thyroid hormone receptors associate with cell growth in human hepatocellular carcinoma
指導教授:林光輝林光輝引用關係
指導教授(外文):K. H. Lin
學位類別:碩士
校院名稱:長庚大學
系所名稱:生物醫學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
論文頁數:80
中文關鍵詞:甲狀腺素甲狀腺素受體長片段非編碼RNASNHG1肝癌
外文關鍵詞:Thyroid hormoneThyroid hormone receptorslncNRASNHG1Hepatocellular carcinoma
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致謝……………………………………………………………………...iii
中文摘要………………………………………………………………...iv
英文摘要………………………………………………………………....v
目錄……………………………………………………………………...vi
圖目錄…………………………………………………………………...ix
表目錄……………………………………………………………………x
第一章 緒論……………………………………...……………………-1-
引言
1.1 甲狀腺素 (thyroid hormone,T3)………………………………-2-
1.2 甲狀腺素受體 (thyroid hormone receptor,TR)……………-3-
1.3 甲狀腺素反應位元………………………………………………-5-
1.4 與甲狀腺素受體作用之調節因子……………………………-6-
1.5 甲狀腺素受體與癌症……………………………………………-7-
1.6 長片段非編碼RNA (long non-coding RNA,lncRNA)………-8-
1.7 Small nucleolar RNA host gene 1 (SNHG1)………………-13-
第二章 研究動機……………………………………………………-15-
第三章 材料與方法…………………………………………………-16-
3.1 細胞培養…………………………………………………………-16-
3.2 甲狀腺素(3, 3’, 5-triiodo-L-thyronine, T3)溶液之配製……-16-
3.3 T3-depleted serum (Td)之配製……………………………-16-
3.4 細胞RNA 萃取……………………………………………………-17-
3.5 反轉錄聚合酶連鎖反應(qRT-PCR)………………………………-18-
3.6 轉染 SNHG1 質體(SNHG1 plasmid transfection)……………-19-
3.7 建立穩定過表現 SNHG1 之肝癌細胞株(SNHG1 stable lines)………-19-
3.8 建立穩定抑制 SNHG1 表現之肝癌細胞株(SNHG1 stable lines)……-20-
3.9 蛋白質萃取……………………………………………………………-21-
3.10 蛋白質定量……………………………………………………………-21-
3.11 西方墨點法(Western blot analysis)……………………………-22-
3.12 細胞週期分析(Cell cycle analysis)……………………………-23-
3.13 細胞增生實驗(Cell proliferation assay)………………………-24-
3.14 細胞遷移實驗(Cell migration assay)………………………-24-
第四章 實驗結果……………………………………………………-25-
4.1 利用Microaray analysis 方式,尋找受T3/TR 調控的lncRNA………-25-
4.2 甲狀腺素抑制lncRNA-SNHG1 的表現……….-25-
4.3 SNHG1 在肝癌患者的腫瘤組織表現量高於其癌旁正常組織………-26-
4.4 甲狀腺素及其受體對SNHG1 調控可能為間接調控…………………-27-
4.5 偵測不同肝癌細胞株之SNHG1 表現量…………………………-27-
4.6 過度表現SNHG1 能夠促進肝癌細胞的增生……………………-28-
4.7 MetacoreTM pathway 分析……………………………………-28-
4.8 SNHG1 抑制p21 的表現量……………………………………-28-
4.9 T3/TR 藉由降低SNHG1 表現進而抑制細胞生長…………-30-
4.10 過度表現SNHG1 能夠促使肝癌細胞的遷移能力………-31-
4.11 T3/TR 負向調節SNHG1-associated snoRNA………-31-
4.12 SNHG1 促進SNHG1-associated snoRNA 的表現………-31-
第五章 討論………………………………………………………….-33-
參考文獻……………………………………………………………...-37-
實驗圖表……………………………………………………………...-49-

圖目錄
圖一、Microarray analysis……………………………………………-49-
圖二、SNHG1 受 T3/TR 的負向調控……………………………...-50-
圖三、肝癌病患檢體中 SNHG1 表現量與存活率分析…………....-51-
圖四、T3/TR 對於 SNHG1 的調控為間接調控…………………..-53-
圖五、偵測 SNHG1 在 HCC cell lines 中內生性表現量………....-54-
圖六、SNHG1 促進肝癌細胞的增生……………………………….-55-
圖七、MetaCoreTM pathway 分析…………………………………...-57-
圖八、SNHG1 抑制 p21 表現,可能藉由影響其蛋白質穩定性....-58-
圖九、T3/TR 藉由降低 SNHG1 表現進而抑制細胞生長………..-60-
圖十、SNHG1 促進肝癌細胞的遷移能力………………………….-62-
圖十一、T3/TR 負向調節 SNHG1-associated snoRNA…………....-64-
圖十二、SNHG1 促進 SNHG1-associated snoRNA 的表現……....-65-

表目錄
表一、Microarray data 交集之結果………………………………....-67-
表二、Clinicopathologic Correlation of SNHG1 Expression in Human
HCCs…………………………………………………………………..-68-
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