| 摘要: | 摘要
結腸癌（colorectal cancer，CRC）是世界上第二大癌症死亡原因，根據侵犯腸壁、淋巴結和轉移的程度，可分為Dukes A至D四個階段，儘管 CRC 的發展可以通過食物營養素和草藥來調節，但是關於荖藤異丁香酚在 CRC 不同階段的控制的訊息傳導路徑的研究相對較少。因此，本論文的總體目標旨在研究異丁香酚調節 CRC 細胞不同階段的生長、遷移和侵襲的訊息路徑。
使用 SW480（Dukes B 期）和 SW620（Dukes C 期）CRC 細胞，我們發現異丁香酚抑制細胞生長，細胞存活率、細胞數量和集落形成減少就是證明。在較小程度上，異丁香酚影響正常 FHC 結腸細胞的生長。進一步的以傷口癒合試驗表示異丁香酚抑制 SW480 和 SW620 細胞的遷移。此外，Boyden chamber assay證實異丁香酚抑制了這兩種 CRC 細胞的侵襲。當檢查訊息路徑時，觀察到異丁香酚會降低 pAkt、CDK2、CDK4、CDK6、細胞週期蛋白 D1、N-鈣粘蛋白和 Snail 蛋白的水平，並增加 pAMPK、pERK、pJNK、pp38 MAPK、ATF3、LC3β 和 E -鈣粘蛋白的表現，也觀察到異丁香酚對細胞週期蛋白 D3、p21 和 p27 蛋白水平的階段依賴性影響，酶譜分析表明異丁香酚抑制 MMP-2 和 MMP-9 蛋白的活性。
使用 AMPK（Compound C）和 p38（SB203580）抑製劑治療可拮抗異丁香酚對 CRC 細胞生長、遷移和侵襲以及 SW480 和 SW620 細胞中大多數訊息蛋白的影響。有趣的是，用 ERK (U0126) 和 JNK (SP600125) 抑製劑治療會增強異丁香酚對 CRC 細胞生長、遷移和侵襲以及大多數訊息蛋白的影響。
這些數據表明，異丁香酚可能通過活化 AMPK 和 p38 MAPK 來抑制 CRC 細胞的生長、遷移和侵襲，並使 CRC 細胞能夠抑制表皮特徵轉變為具有 MMP 依賴性的間質特徵。本論文關於異丁香酚調節 CRC 細胞各個階段活動的訊息機制的結果，將突出其在預防和治療 CRC 方面的可能用途。
;ABSTRACT
Colorectal cancer (CRC) is the second common cause of cancer death in the world and can be divided into four stages from Dukes A to D according to the degrees of invasion of intestinal wall, lymphatic node involvement, and metastasis. Although development of CRC can be regulated by food nutrients and herbal medicines, relatively little information is known about the control of Piper betle isoeugenol in different stages of CRC. Thus, the overall objective of this thesis was designed to study the signal pathways involved in isoeugenol modulations of growth, migration, and invasion in different stages of CRC cells.
Using SW480 (Dukes B stage) and SW620 (Dukes C stage) CRC cells, we found that isoeugenol inhibited cell growth, as evidenced by decreased cell viability, cell number and colony formation. To a much lesser extent, isoeugenol affected the growth of normal FHC colon cells. Further wound healing assay indicated that isoeugenol inhibited the migration of SW480 and SW620 cells. In addition, the Boyden chamber assay confirmed that isoeugenol inhibited the invasion of these two CRC cells. When the signaling pathways were examined, isoeugenol was generally observed to decrease levels of pAkt, CDK2, CDK4, CDK6, cyclin D1, N-cadherin and Snail proteins and increase levels of pAMPK, pERK, pJNK, pp38 MAPK, ATF3, LC3β and E-cadherin proteins. The stage-dependent effects of isoeugenol on levels of cyclin D3, p21, and p27 proteins were also observed. Zymography assay indicated that isoeugenol inhibited the activities of MMP-2 and MMP-9 proteins.
Treatment with the inhibitors of AMPK (compound C) and p38 (SB203580) antagonized the effects of isoeugenol on CRC cell growth, migration, and invasion and most of those signaling proteins in SW480 and SW620 cells. Interestingly, treatment with the inhibitors of ERK (U0126) and JNK (SP600125) generally enhanced the effects of isoeugenol on CRC cell growth, migration, and invasion, and most of those signaling proteins.
These data suggest that isoeugenol may inhibit CRC cell growth, migration and invasion through activations of the AMPK and particular p38 MAPK member, as well as enabling CRC cells to suppress the epithelial features changing to mesenchymal ones with MMP dependency. Results of this thesis for signaling mechanisms of isoeugenol to coordinate the activity of various stages of CRC cells would highlight its possible uses for prevention and cure of CRC. |
