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研究生:吳柏寬
研究生(外文):Bo-Kuan Wu
論文名稱:HBx基因轉殖小鼠之表現型分析
論文名稱(外文):Phenotypic analysis of the HBx transgenic mice
指導教授:蔡亭芬
指導教授(外文):Ting-Fen Tsai
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:遺傳學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:60
中文關鍵詞:基因轉殖小鼠肝癌
外文關鍵詞:HBxHCCtransgenic mice
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根據許多之前臨床以及實驗的研究顯示,HBx在肝癌生成的過程中扮演著舉足輕重的角色,但是HBx與肝癌致病機轉的相關性卻仍不清楚。因此為了解析HBx的生物功能,我們藉由基因轉殖小鼠為研究模型,策略性的應用肝組織啟動子Albumin去調控HBx表現,成功建立了在肝組織中大量表現HBx基因的Alb-HBx基因轉殖小鼠。在肝臟再生的方面,為了更進一步瞭解在肝臟過量表現的HBx是否會影響到肝細胞的細胞週期,我們採用切肝的手術操作去刺激肝臟進行再生。我們首先觀察到Alb-HBx基因轉殖小鼠在手術後的存活率極低,而且肝組織再生的復原情形較差。透過細胞週期相關基因的表現,DNA的複製以及染色體分離等分析,我們推測HBx過量表現會造成肝臟在再生過程中,肝細胞的細胞週期被抑制在G1的階段,導致無法進入正常的細胞週期。此外,從組織病理的分析,我們也發現Alb-HBx基因轉殖小鼠在手術後,其肝臟細胞有許多異常死亡的情形。另一方面,我們也試圖去研究HBx是否會造成肝細胞的病變?初步的實驗結果顯示:從六個月大的Alb-HBx公鼠中,開始觀察到異常肝臟脂肪堆積的現象,而且在十四個月大的Alb-HBx公鼠中,也發現有肝腫瘤的發生,透過組織病理學分析,HBx在小鼠肝臟所引發的腫瘤,相似於人類偽腺體(acinar)特徵的肝癌。系統性肝病變及肝癌發生與HBx基因的相關性則仍待收集更多的實驗樣本來做進一步的分析。
The hepatitis B virus X protein (HBx) has been suggested to play a major role by clinical data and experimental evidences on HBV-induced hepatocellular carcinoma (HCC) development. However, the relationship of HBx and carcinogenesis remains controversial. In order to investigate the physiological effects of HBx, we generated four independent lines of transgenic mice expressing HBx driven by the liver-specific albumin promoter. The effect of HBx over-expression on regenerating liver was studied by two-thirds partial hepatectomy (PHx) strategy which provides an excellent in vivo model for studying hepatocyte cell cycle progression and proliferation. Low survival rate and decreased liver mass recovery of HBx transgenic mice was observed after PHx comparing with wild-type control. Our preliminary data indicated G1 arresting of cell cycle progression in the regenerating liver of the HBx transgenic mice based on analysis of (1) expression profile of cell cycle related genes, (2) BrdU incorporation of DNA synthesis, and (3) mitotic index of regenerating liver. In addition, prolonged mRNA expression of the immediate early genes, i.e. c-myc, c-fos, c-jun and junB, as well as abnormal cell death was detected in the regenerating liver of HBx transgenic mice post-hepatectomy. On the other hand, pathological analysis of the HBx transgenic liver indicated that hepatic steatosis was observed as early as 6 months of age. HCC was developed on 14-month-old transgenic liver with acinar and pseudoglandular histopathological characters of human HCC. The pathological effects of HBx are currently under investigation.
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