- 學位論文
洛神多酚經由調節CD44訊息路徑抑制人類腸癌細胞的轉移
Hibiscus sabdariffa Polyphenol-rich extract (HPE) via regulate CD44 signal pathway to anti-metastatic effect on human colon cancer.
摘要
近年來，用天然的植物萃取出有效的成分，進而預防或抑制癌症的發生已經是目前研究的趨勢。洛神多酚Hibiscus sabdariffa Polyphenol-rich extract (HPE)，洛神花中的功能成分之一，已有許多報告證明洛神花多酚具有促進凋亡、抗氧化活性，降低血脂肪，預防動脈粥狀硬化及防癌等療效，但對於是否對於腸癌腫瘤的轉移造成影響尚未釐清，所以接下來進一步探討洛神多酚是否會抑制腸癌細胞的轉移，並探討其中的作用機制為何。首先我們進行MTT assay，使用DLD-1腸癌細胞株處理HPE，觀察DLD-1細胞的存活率，接著以DLD-1細胞為實驗模式，並以半致死劑量(IC50)的HPE(1、2、3 mg/ml)進行以下的實驗。在DLD-1細胞傷口癒合實驗，我們發現隨著加入HPE劑量越高，細胞爬行的能力愈來越弱；接著在細胞浸潤及轉移的實驗，可以從boyden chamber 及transwell assay的結果觀察到，隨著加入的劑量越高，細胞浸潤及轉移的能力也越來越受到抑制；在Zymography的實驗中觀察了相關蛋白酶MMP及uPA的表現，結果發現HPE對MMP-2及uPA的活性有明顯的抑制效果，並且呈現濃度依賴性。為了釐清其分子機制，我們藉由細胞骨架蛋白、黏附分子及Akt signal pathway的抑制路徑證實，HPE經由調節CD44的訊息路徑有效的抑制DLD-1腸癌細胞的轉移及侵襲作用。根據本篇的結果洛神多酚其具有抗腫瘤轉移的療效，未來可應用於化學預防上成為發展新藥。
並列摘要
In recent years, a natural plant extract an effective ingredient, thereby preventing or inhibiting the occurrence of cancer research has been the current trend. Roselle polyphenols Hibiscus sabdariffa Polyphenol-rich extract (HPE), one of the component in Roselle’s function, our previous studies indicate that HPE can effectively promote apoptosis on gastric cancer and colon cancer cell, the impact on tumor metastasis has not been clarified, so the next one probe into whether HPE inhibit cancer cell metastasis, and to explore the mechanism. We conducted MTT assay, using the DLD-1 colon cancer cells treated with different doses of HPE, observations DLD-1 cell viability, followed by DLD-1 cell as the experimental model, a fine cells induced lethal dose (IC50) of the HPE (1,2,3 mg / ml) for the following experiments. First observation of DLD-1 cells in wound healing experiments, we found that the higher the dose with the addition, DLD-1 cells crawling ability weaker; followed by cell invasion and metastasis experiments by boyden chamber and transwell assay results observed with the addition of the higher dose, DLD-1 cells, invasion and metastasis ability are increasingly inhibited. Zymography we used the experimental observations and related MMP and uPA expression that MMP-2 and uPA activity of HPE in a significant inhibitory effect, and a concentration-dependent manner. In order to clarify the molecular mechanism, we analysis cytoskeletal proteins, adhesion molecules, and the specificity of inhibition of Akt signal pathway to confirmed, HPE via regulating CD44 signaling pathway effectively inhibited DLD-1 colon cancer cell metastasis and invasion effect.