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研究生:林素如
研究生(外文):Su-Ru
論文名稱:相思仔胰蛋白酶抑制劑抑制肺癌細胞侵襲和轉移之研究
論文名稱(外文):Study of the inhibitory effects and mechanisms of Acacia confusa trypsin inhibitor on lung cancer invasion and migration
指導教授:謝易修 博士
指導教授(外文):Yih-Shou Hsieh, Ph.D.
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生化暨生物科技研究所
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:2007
畢業學年度:95
語文別:中文
論文頁數:88
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癌細胞的轉移擴散往往是癌症患者在臨床治療上預後的重要指標,也是主要導致癌症病人死亡的原因。有許多文獻指出在癌症的化學預防上胰蛋白酶抑制劑(trypsin inhibitor)已有許多顯著的生物活性,但是對於其是否會抑制癌症的轉移與其中的詳細機轉至今仍不清楚。在本篇論文中,我們首次證實由相思仔所萃取的胰蛋白酶抑制劑(Acacia confuse trypsin inhibitor ; ACTI)可有效抑制多種肺癌細胞(A549, H1299, and Lewis lung carcinoma (LLC))的細胞侵襲(invasion)、移動(motility)、細胞-基質間的貼附(cell-matrix adhesion)與細胞分泌基質金屬蛋白酶-2、-9 (matrix metalloproteinase (MMP)-2, -9)、尿激酶型纖溶酶原激活物(urokinase-type plasminogen activator; u-PA)的能力。利用西方墨點法,探討其中可能參與的訊息傳遞路徑,證實在A549細胞中ACTI可顯著抑制p-ERK1/2、p-Src、c-Fos與NF-kB的表達,但是對於p-p38與p-Akt則是沒有顯著的影響。此外ACTI也可顯著抑制由12-O-tetradecano-ylphorbol-13-acetate (TPA)所誘導A549細胞細胞的移動能力、MMP-9分泌與p-ERK1/2的表達。最後在動物實驗中,我們利用C57BL/6 老鼠在皮下種植預先處理ACTI的Lewis lung carcinoma,與控制組作比較,會明顯抑制腫瘤的大小。

The invasive ability of tumor cells is a key factor for metastasis and a major cause for treatment failure. Although trypsin inhibitor have been widely recognized to possess several potential as cancer chemopreventive agents, however, limited studies have been available concerning the inhibitory effects of these protein for tumor metastasis. Here, we demonstrated that Acacia confuse trypsin inhibitor (ACTI) could significantly inhibit the invasion, motility, cell-matrix adhesion, secretion of matrix metalloproteinase (MMP)-2, -9, or urokinase-type plasminogen activator (u-PA) of lung cancer cells (A549, H1299, and Lewis lung carcinoma (LLC)). To investigate the possible mechanisms involved in these events, we performed Western blot analysis to find that ACTI inhibited phosphorylation of ERK1/2 and Src, but had no effects on the phosphorylation of p38 and Akt. A treatment with ACTI to A549 cells also inhibited the expression of NF-kB and c-Fos in nuclear extract as shown by Western blot. ACTI inhibited the phosphorylation of ERK1/2, the activity of MMP-9 and cell invasiveness after 12-O-tetradecanoylphorbol-13-acetate (TPA) induction. Finally, these compounds were evidenced by its inhibition on the tumor growth of LLC cells in vivo. Taken together, these findings suggested that ACTI could reduce the invasion and metastasis of lung cancer cells in vivo and in vitro, thereby constituting an adjuvant treatment for metastasis.

目錄
目錄………………………………………………………………1
中英文摘要…………………………………………………………2
縮寫檢索表…………………………………………………………4
壹、緒論……………………………………………………………7
貳、研究動機………………………………………………………26
叁、實驗材料及方法………………………………………………27
肆、實驗結果………………………………………………………41
伍、討論……………………………………………………………47
陸、參考文獻………………………………………………………53
柒、圖表與圖表說明………………………………………………76
捌、附圖表…………………………………………………………86


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