系統識別號 U0026-1907201216534600
論文名稱(中文) 研究Eps8-Ku70交互作用在癌細胞增生中所扮演的角色
論文名稱(英文) Participation of Eps8-Ku70 interaction in cancer cell proliferation
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 100
學期 2
出版年 101
研究生(中文) 許涓藍
研究生(英文) Chuan-Lan Hsu
學號 s26994100
學位類別 碩士
語文別 中文
論文頁數 81頁
口試委員 指導教授-呂增宏
中文關鍵字 Eps8  Ku70  EGFR  STAT3  細胞增生  HeLa cell 
英文關鍵字 Eps8  Ku70  EGFR  Stat3  cell proliferation  HeLa cell 
中文摘要 Ku70影響許多細胞生理功能,像是V(D)J recombination、DNA修復、telomere maintenance和細胞凋亡。然而,近年來Ku70的研究大多都是DNA修復方面的研究,至於在細胞生長分裂的角色則不清楚。近年來的研究發現 (1) 在EGFR signaling被抑制的時候,Ku70會和EGFR、DNA-PK產生複合體; (2) Eps8 (EGF receptor pathway substrate No.8)為EGFR的受質,且大量表現Eps8會促進EGF調控細胞增生作用; (3) Eps8在大腸直腸癌細胞和子宮頸癌細胞會有大量表現的情形,若專一抑制Eps8的表達則可以抑制腫瘤的形成。由於在yeast two-hybrid system中, 我們發現Eps8 和Ku70會有交互作用的產生,因此,我們認為Ku70可能參與Eps8引發的癌細胞形成。首先,我們利用siRNA技術 knockdown Ku70,發現knockdown Ku70會抑制細胞生長和colony 形成,且在Ku70 knockdown的細胞送入Eps8會使此現象有回復的情形。另外,在EGF刺激下,我們發現Ku70 knockdown會使EGFR下游的STAT3磷酸化被抑制,且此現象也會被外送的Eps8回復。進一步我們發現Ku70 knockdown所造成的STAT3活化被抑制是因為Eps8和IRSp53的交互作用被抑制所導致。總結以上,我們發現Ku70 knockdown會造成Eps8和IRSp53交互作用被抑制,進而減少STAT3的活化,導致細胞增生被抑制。
英文摘要 The DNA end-joining protein Ku70 is important in the maintenance of genomic DNA integrity and is involved in many cellular processes, including V(D)J recombination, DNA repair, telomere maintenance and cell death. However, the role of Ku70 in cell proliferation is still vague. Given that EGFR interacts with both DNA-PK and Ku70 when EGFR-mediated signaling is inhibited, Ku70 might participate in EGFR-mediated mitogenesis. In a yeast two-hybrid screening, we have identified Ku70 as an Eps8-interacting protein. In addition, we observed that attenuation of Ku70 in HeLa cells inhibited the cell proliferation and colony formation in soft agar, which could be rescued by ectopically expressed Eps8. Meanwhile, attenuation of Ku70 abolished EGF-mediated Stat3 activation. And, ectopic Eps8 could recover this defect. Finally, Ku70 knockdown decreased the IRSp53-Eps8 interaction in EGF-stimulated HeLa cells. Taken together, our data suggested that Ku70 attenuation reduced the interaction between Eps8 and IRSp53 which resulted in the suppression of EGF receptor-mediated Stat3 activation and the following cell proliferation.
論文目次 中文摘要 1
英文摘要 3
縮寫簡表 7
第一章 緒論 9
第二章 實驗材料與方法 18
第一節 實驗材料 19
第二節 實驗方法 22
第三章 實驗結果 41
第四章 討論 51
第五章 圖表 57
參考文獻 76

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