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系統識別號 U0026-0812200915262431
論文名稱(中文) 分析 AURKC 與 FAZF 蛋白質之間的交互作用與其參與癌化機轉之研究
論文名稱(英文) Characterization of the interaction and function between AURKC and FAZF in tumorigenesis
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 97
學期 2
出版年 98
研究生(中文) 莊凱媛
研究生(英文) Kai-Yuan Chuang
電子信箱 s2696408@mail.ncku.edu.tw
學號 s2696408
學位類別 碩士
語文別 中文
論文頁數 63頁
口試委員 指導教授-王育民
口試委員-許世賢
口試委員-洪良宜
指導教授-張文昌
中文關鍵字 極光激酶 
英文關鍵字 AURKC  FAZF 
學科別分類
中文摘要 極光激酶家族 (Aurora kinase family,AURK) 為調控細胞正常分裂的重要激酶蛋白,在哺乳類動物中 AURK 包含三個成員分別為 AURKA、B、C,其中 AURKC 目前被報導因其序列及結構上與 AURKB 有較高的相似性,因此也與 AURKB 一同參與在細胞染色體及胞質分離的調控機制上。此外,AURKC 在睪丸組織中有較高量的表現,並在精子的生成(spermatogenesis) 調控機制中扮演重要的角色。然而目前對於可受 AURKC 專一性調控的激酶受質所知有限。已知在許多腫瘤組織檢體以及細胞株中都可以發現 AURKC 過度表現的情況,因此推測 AURKC 可能在細胞癌化的機轉上扮演著促進的角色,但其中的致癌機轉仍不清楚。以酵母菌雙雜合篩尋系統,找出與 AURKC 有交互作用的蛋白質- FAZF (Faconi anemia zinc finger protein)。針對 FAZF 進行分析,發現在 HeLa 細胞中大量表現 FAZF 會抑制其細胞繁殖增生以及細胞轉型的能力,而這種抑制的情況可能是由於大量表現 FAZF 後,會誘導細胞停滯在 S/G2/M 等 4N 時期,以致於引發後續的細胞凋亡現象。另一方面,AURKC 蛋白質的活化會大幅降低 FAZF 蛋白質的穩定性,此現象可能影響 FAZF 的抑癌活性。接著分析許多子宮頸癌細胞株,在其中都可以觀察到 AURKC 蛋白質的過度表現伴隨著 FAZF 表現量下降或是不表現的情況,因此本研究推論,FAZF 可能為腫瘤生成抑制蛋白,但由於 AURKC 過度的活化,造成 FAZF 抑制細胞癌化的能力受到限制,因而造成細胞易於癌化 (cell transformation)。
英文摘要 Aurora kinase family (AURK) contains three members, including AURKA, AURKB, and AURKC, and play important roles in the regulation of cell division. The sequence and structure of AURKC show higher similarity with AURKB. Therefore, it was suggested that AURKC may compensate the function of AURKB in regulation of cell mitosis. Comparing the expression of AURKC among various tissues, it shows the highest expression in testis, and is suggested to serve a critical role in spermatogenesis. In contrast to AURKA and AURKB, characterization of AURKC is rather limited, including its substrates. Over-expression of AURKC was observed in many cancer cell lines and cancer specimens, which implied that AURKC may act as an oncogenic protein. However, the AURKC biology in tumorigenesis remains less studied. In this study, a yeast two-hybrid assay was performed and an AURKC-interacting protein, FAZF (Fanconi anemia zinc finger) was identified. Moreover, the interaction of FAZF and AURKC was verified by immuno-precipitation assay. In addition, FAZF was suggested to act as a tumor suppressor by the proliferation and anchorage-independent assays. Its over-expression can induce cell arrest in 4N stages and sub-G1 accumulation in cell cycles. The constitutively-activated AURKC can result in the instability of FAZF. Furthermore, an opposite pattern of protein expression between AURKC and FAZF was observed in cervical cancer cell lines. Taken together, these data suggested that FAZF has the potential to serve as a tumor suppressor, and increase of AURKC activity may impair the stability of FAZF in cervical cancer, which takes an advantage for normal cell be prone to cancer cells.
論文目次 目錄
中文摘要 .....................................................................................1
Abstract .......................................................................................2
誌謝................................................................................................................3
附圖目錄表 .................................................................................7
縮寫指引.......................................................................................................8

第一章 緒論 ..............................................................................10
第一節 極光激酶 (Aurora Kinase)
第二節 Fanconi Anemia Zinc finger protein (FAZF)
第三節 研究動機

第二章 實驗材料及方法 ..........................................................16

第三章 實驗結果 ......................................................................38
第一節 利用酵母菌雙雜系統篩尋與AURKC 的交互作用蛋白質
第二節 FAZF 在哺乳類動物細胞中可與 AURKC 有交互作用
第三節 FAZF 在細胞癌化過程中可能扮演著腫瘤生成抑制的角色
第四節 AURKC 的活化會降低 FAZF 蛋白質的穩定度
第五節 AURKC 過度表現伴隨著 FAZF 蛋白質不表現可能為造成細胞癌化的原因

第四章 討論 ..............................................................................44

第五章 參考文獻 ......................................................................50

附圖 ...........................................................................................56
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