紅茶酒精萃取物抑制人類口腔癌細胞SCC-4之上皮間質轉換機制及體內腫瘤生長能力

中山醫學大學機構典藏 CSMUIR > 醫學院 > 生化微生物免疫研究所 > 博碩士論文 > Item 310902500/10010

Please use this identifier to cite or link to this item: https://ir.csmu.edu.tw:8080/ir/handle/310902500/10010

Title:	紅茶酒精萃取物抑制人類口腔癌細胞SCC-4之上皮間質轉換機制及體內腫瘤生長能力 Black Tea Ethanol Extracts Reverse Epithelial-Mesenchymal Transition of Human Oral Cancer SCC-4 in Vitro And Inhibit Tumor Growth in Vivo
Authors:	陳怡婷 Chen, Yi-Ting
Contributors:	中山醫學大學:生化暨生物科技研究所;陳霈霓
Keywords:	紅茶萃取物;口腔癌細胞;上皮間質轉換機制 Black Tea Extracts;Oral Cancer;Epithelial-Mesenchymal Transition
Date:	2014
Issue Date:	2014-12-10T03:46:38Z (UTC)
Abstract:	腫瘤的轉移是多步驟且複雜的過程，造成臨床上難以治療的原因之一，許多研究觀察，上皮細胞間質化是癌細胞侵襲移動的必要表現，試圖在此過程中，找出抑制或反轉的方法做為治療方針。目前有研究指出，紅茶多酚中許多成分為抗氧化劑，並可抑制腫瘤生成，但這些研究多為說明紅茶多酚如何使癌細胞走向凋亡及其抗氧化作用，較少著重於紅茶萃取物如何減緩癌細胞的侵襲及反轉上皮細胞間質化轉換作用的機制。因此，在我們的研究中，選用人類口腔的惡性鱗狀上皮細胞：SCC-4，以紅茶萃取物處理後，探討紅茶萃取物對於癌細胞表皮細胞間質化及侵襲能力的影響。首先將紅茶茶葉風乾，使用乙醇萃取後，利用不同濃度的紅茶萃取物進行wound healing assay，發現紅茶萃取物有抑制SCC-4移動的能力。接著使用紅茶萃取物處理SCC-4後進行Western blot測定，發現與細胞附著能力有關的蛋白p-paxillin、p-FAK及p-Src表現量下降，與細胞移動有關的蛋白calpain-2表現量也隨之下降，此外，發現SCC-4間質細胞的特徵vimentin會減少，反之上皮的細胞特徵E-cadherin會增加，此外，抑制E-cadherin的轉錄因子snail-1表現量下降，接著利用免疫螢光法再次確認，vimentin表現量明顯下降，綜合上述結果顯示出細胞的侵襲及轉移能力降低。再者，當癌細胞移動時會伴隨著細胞外基質的分解，在gelatine zymography及casein zymography中發現，紅茶萃取物能減少由PMA所誘導增量的MMP-9及u-PA。接著將SCC-4皮下注射進入裸鼠體內，將帶有腫瘤的裸鼠餵食水或紅茶萃取物，發現紅茶萃取物能使腫瘤的體積縮小且重量減輕，而且不具有生物毒性。綜合上述結果，在本篇研究當中證明紅茶萃取物確實能夠透過減少MMPs及u-PA的表現量，來達到反轉人類口腔癌細胞上皮細胞間質化的現象及抑制癌細胞的生長能力。 Tumor metastasis is a complex multistep process and the major cause of cancer death and various treatment strategies have targeted on preventing the occurrence of metastasis. Epithelial to mesenchymal transition (EMT) is critical for the progression, invasion, and metastasis of epithelial tumorgenesis. Various treatment strategies have targeted the prevention of the occurrence of metastasis and reverse EMT. Studies have shown that tea polyphenol, have many anti-oxidant components and could inhibit tumorigenesis, including ovarian cancer and breast cancer. It has been showed that tea polyphenols may have potentially beneficial effects, including anti-metastatic, anti-inflammatory, anti-oxidant and apoptotic effects. However, effects molecular mechanism of black tea in human oral cancer is presently unknown. In this study, anti-metastasis, anti-tumor agents and reversion of EMT were investigated using black tea extracts (BTE), which was extracted by 50% ethanol, on a human malignant oral squamous cell line SCC-4. We demonstrated that BTE could inhibit the migration ability of SCC-4 by wound healing assay. Immunoblot was performed to find that BET could inhibit the proteins associated with cell adhesion ability such as p-paxillin、p-FAK and p-Src. BTE was down-regulation of calpain-2, a proteins associated with cell migrasion. Otherwise, mesenchymal marker vimentin expression was decreased. BTE could induce up-regulation of epithelial markers such as E-cadherin. Expression of transcription factor Snail-1 was decrease. Recomfirmed by immunofluorescence, Vimentin expression was significantly decreased. Found in gelatine zymography and casein zymography that BTE inhibits PMA-induced MMP-9 and u-Paexpression in SCC-4. BTE also inhibit the tumor growth of SCC-4 cells via cancer cell xenografted nude mice. We evidence, that BTE could reverse EMT and inhibit cancer growth ability in human oral cancer cells.
URI:	https://ir.csmu.edu.tw:8080/ir/handle/310902500/10010
Appears in Collections:	[生化微生物免疫研究所] 博碩士論文

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