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研究生:黃仁星
研究生(外文):Jen-Hsin Huang
論文名稱:蛋白質酪胺酸激脢抑制劑—類黃鹼素,抑制A431癌細胞轉移、生長之機制探討
論文名稱(外文):Flavonoids, tyrosine kinase inhibitors, effect on A431 cell growth and metastasis-associated properties
指導教授:李明亭
指導教授(外文):Ming-Ting Lee
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:生化科學研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:1999
畢業學年度:87
語文別:中文
論文頁數:53
中文關鍵詞:類黃鹼素上皮生長因子受體蛋白質酪胺酸激脢轉移
外文關鍵詞:flavonoidepidermal growth factor receptorprotein tyrosine kinasemetastasis
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類黃鹼素 (Flavonoids) 廣泛地存在於高等植物之中,平均而言,常人每日從飲食中攝取的類黃鹼素可高達1公克左右。目前類黃鹼素被發現具有的生物功能中包括:抑制癌細胞轉移、抑制癌細胞週邊血管增生、及誘導細胞凋亡(apoptosis)發生等。此外,關於類黃鹼素在抗癌活性的研究方面,發現類黃鹼素是藉由抑制調控細胞生長、分化的特定酵素,來達到對抗癌細胞生長的目的,而這些特定的酵素多為蛋白質酪胺酸激脢。
本實驗的研究主題為A431 癌細胞,為一高度表現EGF receptor的人類上皮細胞癌;經實驗測試八種不同的類黃鹼素發現luteolin在抑制A431癌細胞株生長方面,有相當明顯的效果(IC50 = 9 μM);在20 μM的濃度下,luteolin及quercetin抑制EGFR autophospho-rylation程度超過75 %以上,且luteolin及quercetin是透過與ATP的競爭達到抑制EGFR磷酸化,在濃度20μM下,Ki值分別為6.85及9.70μM。此外,實驗結果指出,luteolin 、quercetin、genistein確實可誘導細胞進行細胞凋亡,有DNA片斷梯度、染色體斷裂的現象發生;再觀察細胞培養液中MMP-2及MMP-9活性時發現luteolin 、quercetin、genistein和d-catechin可抑制MMP-2及MMP-9的活性,且抑制MMPs活性的機制,可能是透過抑制細胞分泌MMPs,不是因抑制細胞生長來達到抑制MMPs活性的效果,作為癌細胞是否發生轉移的依據。
綜合以上結果,我們推論luteolin 及quercetin,未來有希望成為對抗癌症的化合物 (藥物) 之一。

The flavonoids are present in almost all-higher plants and their consumption in the normal diet amounts to 1 g or more per day. Flavonoids display a wide range of pharmacodynamic properties including anti-inflammatory, anti-mutagenic, anti-metastatic, anti-cacinogenic and anti-cancer effects. Resent studies have demonstrated that, depending on their structure, flavonoids are inhibitors of several kinases involved in signal transduction, mainly tyrosine kinases.
In this study, we evaluated the effects of eight flavonoids on EGFR-overexpressed human epidermoid carcinoma cells A431 proliferation, EGFR phosphorylation, and matrix metalloproteinases (MMP) secretion. Of the flavonoids examined, luteolin significantly inhibited A431 cell proliferation with IC50 value of 9 μM. At a concentration of 20μM, both luteolin and quercetin markedly decreased the level of phosphorylation of EGFR. We estimated that luteolin and quercetin can inhibit the EGFR kinase activity by competing with ATP, and Ki values of 6.85μM and 9.70μM for 20μM, respectively. Our data show that luteolin, quercetin and genistein could induce apoptosis in A431 cell. We also determined that luteolin, quercetin, genistein and d-catechin could suppress the activity of MMP-2 and MMP-9 in A431 cells at low concentration. Treatment with flavonoids may inhibit secretion of these MMPs from tumor cells, and thereby reduce the potential for metastasis.
This study supports the idea that luteolin and quercetin may have potential as anti-cancer and anti-metastasis agents.

目錄i
圖表目錄ii
縮寫表iii
中文摘要iv
英文摘要v
第一章 前言 1
The Epidermal Growth Factor receptor (EGFR) 3
Flavonoids 7
第二章 材料與方法12
第三章 結果21
類黃鹼素對A431癌細胞株生長測試21
Luteolin、quercetin、genistein和d-catechin抑制EGFR auto- 及 transphosphorylation 活性測試22
Luteolin、quercetin抑制EGFR酵素特性測試23
Luteolin、quercetin、genistein和d-catechin抑制A431癌細胞株生長機制討論24
Luteolin、quercetin、genistein和d-catechin在A431癌細胞株抑制MMPs活性測試25
第四章 討論43
Luteolin、quercetin及genistein抑制A431癌細胞株生長43
Luteolin、quercetin和genistein抑制EGFR auto- 及transphosphorylation44
Luteolin、quercetin及genistein誘導A431癌細胞株之細胞凋亡 45
Luteolin、quercetin、genistein和d-catechin在A431癌細胞株抑制MMPs活性46
結論48
參考文獻49

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