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系統識別號 U0026-2608201402120700
論文名稱(中文) 轉錄活化Pentraxin3基因表現與表皮生長因子誘導頭頸癌細胞轉移具有相關性
論文名稱(英文) Transcriptional activation of pentraxin-3 gene expression is associated with EGF-induced head and neck cancer cell metastasis
校院名稱 成功大學
系所名稱(中) 生物資訊與訊息傳遞研究所
系所名稱(英) Insitute of Bioinformatics and Biosignal Transduction
學年度 102
學期 2
出版年 103
研究生(中文) 吳碩倫
研究生(英文) Shuo-Lun Wu
學號 z26011071
學位類別 碩士
語文別 中文
論文頁數 72頁
口試委員 指導教授-陳炳焜
口試委員-王育民
口試委員-張偉嶠
中文關鍵字 頭頸癌  表皮生長因子  PTX3  癌症轉移 
英文關鍵字 Head and neck cancer  epidermal growth factor  PTX3  metastasis 
學科別分類
中文摘要 在頭頸癌中,表皮生長因子受器的過度表現和促發炎之細胞素的產生已充分釐清且與癌細胞侵入以及轉移有所關連。然而,表皮生長因子受器與細胞素調控頭頸癌細胞轉移之分子機制所知仍然相當缺乏。在本篇研究,我們確認PTX3是表皮生長因子調控頭頸癌轉移中的促進因子。藉由比較分析資料庫中正常與頭頸癌病人組織之間或病人原位癌組織與癌轉移組織之間PTX3表現,指出PTX3在惡性或癌轉移組織表現高於正常組織。我們發現表皮生長因子透過轉錄活化PTX3,藉由活化轉錄因子NF-κB和c-Jun結合啟動子AP-1/NF-κB結合位。探討表皮生長因子其下游訊息傳導路徑,發現PI3K/AKT與NF-κB訊息路徑對PTX3的活化是重要的。此外,分別抑制NF-κB和c-Jun狀態下,抑制表皮生長因子調控PTX3表現。表皮生長因子刺激PTX3分泌,促進頭頸癌細胞爬行、局部侵入、與內皮細胞的黏附和遠端傳播。表皮生長因子活化Fibronectin、MMP9與E-cadherin的抑制,在shPTX3細胞中被抑制。以上結果指出表皮生長因子調控PTX3促進頭頸癌細胞的轉移,可能透過PTX3自迴分泌調控促進轉移分子例如Fibronectin與MMP9。PTX3的活化也許反映表皮生長因子促進頭頸癌細胞的轉移與發炎反應有所關連。
英文摘要 SUMMARY

Overexpression of epidermal growth factor receptor (EGFR) and production of proinflammatroy cytokines are well clarified in head and neck squamous cell carcinoma (HNSCC) and correlates with invasion and metastasis. However, molecular mechanisms by which EGFR-mediated cytokines regulates metastasis of HNSCC remain unclear. Here, we identified that PTX3 is a metastasis-promoting factor that mediate EGF-induced HNSCC metastasis. Analysis of PTX3 expression between normal and malignant or metastatic tissues from HNSCC patients by using published datasets, our results indicated that expression level of PTX3 in malignant tissues was higher than in normal part in HNSCC patients. In addition, we found that EGF induced transcriptional activation of PTX3 by activating the binding of c-Jun and NF-κB factors to AP-1/NF-κB binding site of the PTX3 promoter. PI3K/AKT and NF-κB were essential for the induction of PTX3. EGF-induced PTX3 expression was significantly inhibited in c-Jun and NF-κB knockdown cells. Furthermore, we identify that NF-κB is upstream regulator to c-Jun in EGF signaling pathway. EGF stimulated the secretion of PTX3 from cancer cell, resulting in enhancing cell migration, local invasion, attachment with endothelium and distal dissemination. Effects of EGF on the induction of fibronectin and MMP9 expression, and inhibition of E-cadherin were abolished in PTX3 knockdown cells. These findings reveal the mechanism that autocrine production of EGF-induce PTX3-regulated HNSCC metastasis was through enhancing metastatic molecules, such as fibronectin and MMP9 expression. Induction of PTX3 possibly reflecting the EGFR-caused HNSCC metastasis associated with inflammation.
論文目次 目錄

摘要--------------------i
Abstract---------------ii
致謝--------------------v
目錄-------------------vi
圖目錄---------------viii
附圖目錄---------------ix
第一章 緒論-----------------------------------1
第一節 頭頸癌簡介--------------------------1
第二節 表皮生長因子對癌細胞轉移之影響--------1
第三節 Pentraxin 3(PTX3)簡介-------------2
(一) PTX3之起源--------------------2
(二) PTX3在組織中分布、調控與生理功能-------3
第四節 Pentraxin 3(PTX3)在癌症中的角色----------5
(一) 軟組織脂肪肉瘤(Soft tissue liposarcoma)--5
(二) 肺癌(Lung cancer)-------------------5
(三) 乳癌(Breast cancer)-----------------6
(四) 前列腺癌(Prostate cancer)-----------6
(五) 黑色素瘤(Melanoma)-----------------6
第五節 研究動機----------------------------------7
第二章 材料與方法------------------------------------8
第三章 實驗結果----------------------------------------19
I. 探討表皮生長因子是否誘導頭頸癌細胞表現PTX3-------19
(一) 探討PTX3於頭頸癌的表現情形-------------------19
(二) 探討表皮生長因子是否刺激頭頸癌細胞表現PTX3----19
II. 探討表皮生長因子活化PTX3是否透過NF-κB訊息傳導路徑---20
(一) 探討NF-κB是否參與表皮生長因子誘導PTX3的表現---20
III. 探討NF-κB在PTX3啟動子上的轉錄因子結合位---------21
(一) 探討表皮生長因子是否誘導c-Jun與NF-κB是否結合PTX3啟
動子的AP-1轉錄因子結合位-----------------------21
(二) 探討表皮生長因子誘導NF-κB是否結合AP-1/NF-κB位點與活
化PTX3啟動子之區間----------------------------22
(三) 探討表皮生長因子活化NF-κB如何調控PTX3表現-------23
IV. 探討表皮生長因子調控PTX3表現在頭頸癌細胞中的功能----24
(一) 探討表皮生長因子活化PTX3對頭頸癌細胞的爬行能力之影
響--------------------------------------------24
(二) 探討表皮生長因子活化PTX3對頭頸癌細胞的侵入能力之影
響--25
(三) 探討表皮生長因子活化PTX3對頭頸癌細胞的轉移能力之影
響--25
V. 探討表皮生長因子調控PTX3參與頭頸癌細胞轉移機制-----26
(一) 探討表皮生長因子透過PTX3活化的癌症轉移相關之基因--26
(二) 探討表皮生長因子透過PTX3活化癌細胞侵入相關之基因--27
第四章 討論----------------------------------------------29
第一節、 表皮生長因子促進頭頸癌細胞活化PTX3表現---------29
第二節、 表皮生長因子調控PTX3表現之機制----------------30
第三節、 表皮生長因子活化PTX3促使頭頸癌細胞轉移---------31
第四節、探討癌細胞自迴分泌之PTX3相互作用之受器----------33
第五節、未來研究方向----------------------------------34
第六節、 總結----------------------------------------35
參考文獻-------------------------------------------------37


圖目錄

圖1. PTX3過度表現於在人類頭頸癌-----------------------45
圖2. 表皮生長因子活化頭頸癌細胞珠表現PTX3--------------47
圖3. NF-κB參與表皮生長因子誘導PTX3的表現--------------48
圖4. 表皮生長因子促進c-Jun與p65結合PTX3啟動子之AP-1/NF-κB轉錄因子結合位-------------------------------------------51
圖5. 表皮生長因子活化頭頸癌細胞爬行能力在致弱PTX3細胞中被抑制-------------------------------------------------------55
圖6. 表皮生長因子活化頭頸癌細胞侵入能力在致弱PTX3細胞中被抑制-------------------------------------------------------56
圖7. 表皮生長因子誘導癌細胞遠端傳播在致弱PTX3細胞中被抑制---59
圖8. 表皮生長因子誘導PTX3活化fibronectin且頭頸癌細胞侵入能力在致弱fibronectin細胞被抑制--------------------------------61
圖9. 表皮生長因子在頭頸癌細胞誘導PTX3活化MMP9表現----------65


附圖目錄

附圖1: 介白素-1β藉由NF-κB活化PTX3啟動子並活化PTX3表現---------66
附圖2: 表皮生長因子活化頭頸癌細胞黏附能力在siPTX3細胞中被抑制-----67
附圖3: 表皮生長因子活化頭頸癌細胞黏附能力在siPTX3細胞中被抑制 ----68
附圖4: 表皮生長因子活化頭頸癌細胞侵入能力在shFibronectin細胞中被抑制-------------------------------------------------------------------69
附圖5: CD44穿膜與胞內區域在PTX3致弱細胞中抑制表現---------------70
附圖6: PTX3與C反應蛋白和血清澱粉蛋白胺基酸序列之差異------------71
附圖7: Gefitinib在膠質母細胞瘤中抑制PTX3表現---------------------72
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