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研究生:蔡志鵬
研究生(外文):Jhih-PengTsai
論文名稱:探討由表皮生長因子促進PTX3表現之機轉及在癌細胞中所扮演的角色
論文名稱(外文):The mechanism and functional role of EGF-induced PTX3 expression in the tumor cells
指導教授:陳炳焜
指導教授(外文):Ben-Kuen Chen
學位類別:碩士
校院名稱:國立成功大學
系所名稱:生物資訊與訊息傳遞研究所
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:2012
畢業學年度:100
語文別:中文
論文頁數:59
中文關鍵詞:表皮生長因子癌細胞
外文關鍵詞:PTX3EGF
相關次數:
  • 被引用被引用:1
  • 點閱點閱:246
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  • 下載下載:8
  • 收藏至我的研究室書目清單書目收藏:0
PTX3屬於Pentraxin的一員,能藉由前發炎因子如interleukin-1β (IL-1β), tumor necrosis factor-α (TNFα)以及lipopolysaccharide (LPS)等促進表現,調控的生理功能包含免疫、基質沉積、血管新生、發炎以及女性生殖方面。先前研究顯示,PTX3過量表現於軟組織脂肪肉瘤中;而於前列腺癌中,PTX3的表現與EGFR呈現正相關。然而,癌細胞中EGF如何調控PTX3,其機制仍是未知。因此本篇的研究目標為探討癌細胞中,EGF如何調控PTX3,以及EGF誘導產生的PTX3,於癌細胞中的功能為何。首先使用即時聚合酶連鎖反應、西方墨點法以及酵素連結免疫吸附分析法,證實EGF能活化癌細胞產生PTX3。藉由使用Actinomycin D抑制基因轉錄實驗,以及使用PTX3報導基因載體進行報導基因實驗,研究EGF透過活化基因轉錄或穩定PTX3 mRNA,使PTX3表現增加。此外,將EGF訊息相關基因致弱,以及使用相關抑制劑抑制基因表現後,發現EGF透過EGFR下游路徑活化PTX3產生。使用功能性實驗,發現EGF促進癌細胞的生理功能,PTX3參與其中。總結此篇研究,EGF促進癌細胞表現PTX3,並且調控癌細胞的生理功能。
PTX3 belongs to the pentraxin superfamily, is induced by pro-inflammatory factors such as interleukin-1β (IL-1β), tumor necrosis factor-α (TNFα), and lipopolysaccharide (LPS), and is involved in immunity, matrix remodeling, angiogenesis, fertility and apoptosis. Previous reports have shown that overexpression of PTX3 was found in the soft tissue liposarcoma and up-regulation of PTX3 had highly correlation with overexpression of epidermal growth factor receptor (EGFR) in the prostate cancer. However, the mechanism involved in EGF-regulated PTX3 expression is not clear. The aim of this study is to characterize whether EGF can induce PTX3 expression and to investigate the functional role of PTX3 in the tumor microenvironment. In the present study, we used RT-PCR, western blotting and ELISA to clarify that EGF-induced the expression of PTX3 was in a time-dependent manner in cancer cells. We used actinomycin D treatment and reporter assay to clarify EGF induces the expression of PTX3 through transcriptional activation or stabilizing mRNA. We silenced genes that are downstream of EGFR-signaling pathway by lentiviral vector-derived shRNAs and found that EGF induced the expression of PTX3 through EGFR-related downstream pathways. In functional assay, PTX3 is involved in EGF-regulated cell function. These results suggested that EGF-induced PTX3 expression was through EGFR pathway, resulting in the regulation of cell function.
摘要 -----------------------------------------------------i
Abstract-------------------------------------------------ii
誌謝-----------------------------------------------------iii
目錄-----------------------------------------------------iv
表目錄----------------------------------------------------vi
圖目錄---------------------------------------------------vii
附錄----------------------------------------------------viii
第一章 緒論-------------------------------------------------1
第一節 Pentraxin 3 (PTX3) 的起源與結構--------------------1
第二節 Pentraxin 3 (PTX3) 的組織分與調控因子---------------1
第三節 Pentraxin 3 (PTX3) 的生理功能---------------------3
第四節 Pentraxin 3 (PTX3) 與癌症形成之關係----------------6
研究動機------------------------------------------------8
第二章 實驗材料與方法-----------------------------------------9
第三章 實驗結果---------------------------------------------16
第一節 EGF誘導PTX3表現於癌細胞中--------------------------16
(一) EGF活化癌細胞表現PTX3--------------------------16
(二) EGF活化PTX3之基因與蛋白表現---------------------17
第二節 探討EGF活化PTX3是透過何種機制----------------------17
(一) 探討EGF是否調控PTX3 mRNA的穩定度----------------17
(二) 探討EGF是否調控PTX3的基因轉錄--------------------18
(三) 探討EGF調控PTX3啟動子的區域---------------------18
(四) 探討PTX3啟動區轉錄因子結合位對於EGF活化啟動子的角色-19
(五) 篩選EGF調控PTX3啟動子的其他可能位置---------=-----20
(六) 探討EGF活化EGFR訊息的何種路徑影響PTX3的表現-------21
第三節 探討EGF活化PTX3於癌細胞中扮演的功能-----------------21
(一) 挑選穩定致弱PTX3的癌細胞株---------------------22
(二) PTX3參與在EGF調控的生理功能--------------------22
第四章 討論------------------------------------------------23
第一節 EGF促進癌細胞表現PTX3------------------------------23
第二節 EGF調控PTX3的機制--------------------------------24
第三節 PTX3參與在EGF影響的生理功能------------------------25
參考文獻---------------------------------------------------28

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