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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李明亭 | |
dc.contributor.author | Chun-Yu Lin | en |
dc.contributor.author | 林俊宇 | zh_TW |
dc.date.accessioned | 2021-06-13T08:07:38Z | - |
dc.date.available | 2006-07-27 | |
dc.date.copyright | 2005-07-27 | |
dc.date.issued | 2005 | |
dc.date.submitted | 2005-07-21 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36608 | - |
dc.description.abstract | 類黃鹼素,主要是一群由polyphenolic物質所組成的benzo-γ-pyrone(phenylchromone)衍生物 。而類黃鹼素又可分為以下幾類:flavonols、flavans、proanthocyanidins、anthocyanidins、flavanones、flavones、isoflavones、neoflavonoids,而這些類黃鹼素廣泛的存在於水果、穀類、豆科植物、蔬菜類、堅果類、種子、草本植物、莖、花、茶、可可亞、啤酒及葡萄酒當中。根據已發表的研究指出,食用性的類黃鹼素之生物性功用包含有抗發炎、抗過敏、抗微生物、抗病毒、保護肝臟、抗凝血、心血管保護、微血管強化、抗糖尿病、抗癌以及抗癌細胞新生之作用。
本文的主要研究目的在於觀察類黃鹼素對於前列腺癌細胞的抑癌作用之探討。我們利用in vitro chemo-invasion assay的方式,成功自Du145-P母代癌細胞中篩選出分泌較多MMPs之Du145-I、Du145-II、Du145-III子代細胞。並且利用類黃鹼素Luteolin及Quercetin探討其對Du145-P及Du145-III之抑制作用。結果顯示,Luteolin及Quercetin會抑制Du145-P及Du145-III之生長,使其外型發生變化,並且誘導apoptosis的現象發生,抑制兩細胞株MMPs之釋出,抑制其轉移以及誘導web forming之能力。而EGF則與Luteolin及Quercetin有相反之效果,其能促進癌細胞之生長,促進MMPs之釋出,促進癌細胞的轉移以及web forming,同時我們比較了Du145-P與Du145-III間之差異,發現兩細胞株不僅外形上有差異,Du145-P較容易黏附在一起,而Du145-III則是較為分散,並且細胞外觀較長也較尖;而在MMPs之釋放方面,Du145-III之釋放量較Du145-P高,轉移以及web forming之能力亦高出許多。 另外我們也觀察到另一株前列腺癌細胞株PC-3,其MMPs之釋放種類亦與Du145類似,因此我們正嘗試篩選不同前列腺癌細胞PC-3、22RV1及LNCAP之第三代子代,觀察其MMPs釋放之種類,希望藉由與不同器官、組織的腫瘤細胞相比較,鑑定出特殊的organ specific MMPs,並且發展出一套可用來診斷前列腺癌的系統,也或許可提供作為新藥開發的線索。 另一方面,我們成功地篩選到能激發出紅色及綠色螢光的Du145-P及Du145-III,在未來將努力在animal model上建立一套觀察前列腺癌細胞轉移路徑的系統,以及觀察其所轉移之特異性組織與器官。另外也期望能在in vivo的系統下,實際去比較Du145-P及Du145-III之入侵能力,進一步佐證在in vitro下所得到的實驗結果。 | zh_TW |
dc.description.abstract | The flavonoids, which are primary benzo-γ- pyrone ( phenylchromone) derivatives, comprise a massive group of polyphenolic compounds. The immensely diverse group broadly comprises distinct classes such as flavonols, flavans and proanthocyanidin, anthocyanidinsm flavanones, flavones, isoflavones and neoflavonoids. They are universally present in fruits, cereals, legumes, vegetables, nuts, seeds, herbs, spices, stems, flowers as well as in beverages such as tea, cocoa, beer and wine. As components of edible plants and plant foodstuffs they constiturte an integral part of the human diet. The documented biological effects of dietary flavonoids include anti-flammatory, antiallergic antimicrobial, hepatoprotective, antiviral, antithrombotic, cardioprotective, capillary strengthening, antidiabetic, anticarcinogenic and antineoplastic effects, among others.
In this thesis, we established the relatively higher invasive potential tumor cell line: Du145-I, II and III from Du145-P (parental) using Boyden chamber invasion assay. The Du145-III secreted higher amount of MMPs, higher migration and angiogenic ability as compared to Du145-P. According to the amount of MMPs secretion, migration and angiogenic ability, we concluded that Du145-III had relative higher invasive potential than that of Du145-P. Luteolin and Quercetin were documented as protein kinase inhibitor, they inhibit the activity of protein kinase by blocking the ATP binding site in catalytic unit. In this study, I tested the anti-tumor effect of Luteolin and Quercetin on Du145-P and Du145-III cells. Luteolin and Quercetin efficiently inhibit the growth of Du145-P and Du145-III cells. In addition, Luteolin and Quercetin both exert the inhibitory effect of MMPs secretion, migration and angiogenesis of Du145-P and Du145-III cells. In a separate study, using PC-3 prostate tumor cell line, we observed that the profile of MMPs secretion is similar to that of Du145. Therefore, we intend to investigate the profile of other prostate tumor cell lines, including 22RV1 and LNCAP, in order to confirm those MMPs are organ specific. We also established the prostate tumor cell lines Du145-P and Du145-III that could excite the red and green fluorescence. In the future, we intend to establish the animal model to observe the metastasis pathway of prostate tumor cell line by using the Du145-P and Du145-III that could excite the red and green fluorescence. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T08:07:38Z (GMT). No. of bitstreams: 1 ntu-94-R92b46019-1.pdf: 8061686 bytes, checksum: acb24bf67f1820068ed3ea555cb926ab (MD5) Previous issue date: 2005 | en |
dc.description.tableofcontents | 目錄
Abstract 1 中文摘要 2 第一章、前言 4 一、酪胺酸磷酸化(tyrosine phosphorylation)與癌症的關係 4 二、酪胺酸激酶(protein tyrosine kinase)及酪胺酸去磷酸酶(protein tyrosine phosphatase)之簡介 5 三、表皮生長因子受體(Epidermal growth factor receptor,EGFR)之簡介 6 四、Focal adhesion kinase(FAK)之簡介 8 五、Matrix metalloproteinases(MMPs)之簡介 9 六、類黃鹼素(Flavonoids)之簡介 10 第二章、材料與方法 13 材料: 13 方法: 13 一、細胞培養(Cell culture) 13 二、細胞之解凍與冷藏 14 三、較具侵入性(invasion potential)癌細胞之篩選 14 四、細胞生長曲線(Cell growth curve experiment) 15 五、結晶紫染色法(Crystal violet stainig) 15 六、細胞蛋白質萃取(Preparation of cell lysate) 15 七、蛋白質濃度測定(Determination of protein concentration) 16 八、SDS聚丙醯胺凝膠電泳分析(SDS polyacrylamine gel electrophoresis,SDS-PAGE) 16 九、西方墨點法分析(Western blotting) 17 十、流式細胞儀分析(Flow cytometry) 18 十一、DNA片段分析(Analysis of DNA fragmentation) 18 十二、梯度電泳分析(Gradient gel electrophoresis) 19 十三、Gelatin zymography 19 十四、Wound healing assay 20 十五、In vitro capillary formation assay 21 第三章、結果 22 壹、Du145-P與Du145-III之比較 22 一、Du145-P及Du145-III生長速率之比較 22 二、Du145-III所釋放之MMPs明顯高於Du145-P 23 三、Du145-III之轉移能力(migration ability)明顯優於Du145-P 23 四、Du145-III之web forming能力優於Du145-P 23 貳、類黃鹼素Luteolin及Quercetin對Du145-P及Du145-III之影響 24 一、Luteolin及Quercetin改變Du145-P及Du145-III之外形 24 二、Luteolin及Quercetin抑制Du145-P及Du145-III之生長 25 三、Luteolin及Quercetin具有誘導Du145-P及Du145-III癌細胞產生程式凋亡(apoptosis)之趨勢 25 四、Luteolin及Quercetin抑制Du145-P及Du145-III之MMP9的釋放,EGF則促進MMP9之釋放 26 五、Luteolin及Quercetin抑制Du145-P及Du145-III之migration,而EGF則促進其migration 26 六、Luteolin及Quercetin抑制Du145-P及Du145-III之web forming能力,而EGF則促進web forming 27 第四章、討論 48 第五章、結論 55 參考文獻 56 圖表目錄 圖一、In vitro chemo-invasion assay…………………………………………………28 圖二、Du145-P與Du145-III間型態上之差異………………………………………29 圖三、Du145-P及Du145-III生長曲線之比較………………………………………30 圖四、Du145-P與Du145-III前列腺癌細胞在EGF處理後MMPs釋放之多寡與差異之比較……………………………………………………………………..31 圖五、利用Wound-healing assay之方式檢測Du145-P與Du145-III之轉移(migration)能力……………………………………………………………32 圖六、由in vitro capillary assay觀察Du145-P與Du145-III之web forming能力……………………………………………………………………………..33 圖七、Luteolin與Quercetin對Du145-P及Du145-P型態上之影響………………..35 圖八、Luteolinu及Qucertin對Du145-P及Du145-III癌細胞生長之影響…………37 圖九、以流式細胞儀(Flow cytometry)觀察Luteolin及Quercetin對Du145-P與Du145-III癌細胞之影響…………………………………………………….39 圖十、EGF、Luteolin及Quercetin對Du145-P及Du145-III癌細胞之DNA完整性(DNA integrity)之影響…………………………………………………...40 圖十一、Luteolin、Quercetin及EGF對Du145-P及Du145-III分泌釋出MMPs之影響…………………………………………………………………………..41 圖十二、Luteolin及Quercetin及EGF對Du145-P及Du145-III癌細胞之轉移(migration)之影響…………………………………………………………46 圖十三、Luteolin、Quercetin及EGF對Du145-P及Du145-III之web forming能力之影響……………………………………………………………………..47 附錄一、Protein tyrosine phosphatase是protein tyrosine kinase之負調控因子……66 附錄二、EGF Receptor 之結構圖…………………………………………………...67 附錄三、Organization of the domains of focal adhesion kinase……………………...68 附錄四、Proposed interactions among the proteins involved in integrin signaling…..69 附錄五、Structure of human matrix metalloproteinases……………………………...70 附錄六、人類細胞中MMPs之特異性受質、染色體位置及結構組成.........................71 附錄七、the matrix metalloproteinase family…………………………......................73 附錄八、MMPs之蛋白質結構………………………………………………………74 附錄九、Functions of MMPs in cancer progression…………………………………75 | |
dc.language.iso | zh-TW | |
dc.title | 類黃鹼素對前列腺癌細胞母代Du145-P及篩選出較有入侵性之子代Du145-III抗癌作用之探討 | zh_TW |
dc.title | The anti-tumor effect of flavonoids on Du145 parental cells and highly invasive potential Du145-III. | en |
dc.type | Thesis | |
dc.date.schoolyear | 93-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張震東,黃銓珍,陳宏文,孟仔青 | |
dc.subject.keyword | 類黃鹼素,前列腺癌,轉移,入侵, | zh_TW |
dc.subject.keyword | flavonoid,MMPs,invasion,metastasis,Du145, | en |
dc.relation.page | 75 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2005-07-21 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
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