檳榔嚼塊萃取液對基質金屬蛋白酶表現的影響

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Title:	檳榔嚼塊萃取液對基質金屬蛋白酶表現的影響 Effects of areca quid extract on the expressions of matrix metalloproteinases
Authors:	陳瑮琳 Li-Lin Chen
Contributors:	林美惠;劉永超嘉南藥理科技大學：生物科技研究所
Keywords:	檳榔基質金屬蛋白酶口腔鱗狀細胞上皮癌 oral squamous cell carcinoma OECM-1 matrix metalloproteinase areca
Date:	2005
Issue Date:	2008-10-31 16:15:18 (UTC+8)
Abstract:	在台灣，嚼食檳榔和口腔癌有很密切的相關性。基質金屬蛋白酶 (matrix metalloproteinases, MMPs) 能夠降解胞外基質 (extracellular matrix, ECM) 和基底膜，在釵h上皮癌症的發展以及生成，它們扮演一個很重要的角色，其中也包括口腔鱗狀細胞上皮癌 (oral squamous cell carcinoma, OSCC)。從有嚼食檳榔習慣的病人中，我們發現有數種MMPs在其口腔腫瘤樣本中有很高的表現率。我們預期檳榔嚼塊萃取液 (Areca quid extract, AQE) 有刺激這些MMPs表現的潛力，間接促進腫瘤進行侵犯和轉移作用；在本研究中以口腔鱗狀細胞上皮癌的細胞株OECM-1當作實驗的模型，並以西方墨點分析法、zymograph或反轉錄聚合酶連鎖反應 (reverse transcription-polymerase chain reaction, RT-PCR) 來檢測MMP-2, -8 和 -28的蛋白質及mRNA經AQE刺激後之變化情形。結果顯示短時間 (10分鐘) AQE的處理 (short-term AQE treatment, SAT) 和長時間 (24小時) 的AQE處理 (long-term AQE treatment, LAT) 都會誘導MMP-2, -8 和 -28蛋白質合成與分泌。藉由使用兩種訊息傳導分子的抑制劑，PI3K (Phosphatidylinositol 3-kinase) 抑制劑及MEK (Mitogen-activated protein kinase kinase) 抑制劑，我們也觀察到SAT與LAT對不同MMPs的誘導作用對PI3與MEK的依賴性並不盡相同；最後由半定量的RT-PCR方法偵測出MMP-2與MMP-28之mRNA會因LAT而增加。 In Taiwan, areca quid (AQ) chewing shows a strong correlation to the incidence of oral cancer. Matrix metalloproteinases can degrade extracellular matrix and basement membrane, and play an important role in the development and progression of various carcinomas, including oral squamous cell carcinoma (OSCC). We have found that several MMPs are commonly expressed in oral tumor specimens from patients with AQ consuming history. It was suspected that areca quid extract (AQE) may have the potential to stimulate the expression of these MMPs, which in turn facilitates tumor invasion and metastasis. An OSCC cell line, OECM-1, was utilized as the experimental model in this study. Western blot analysis, zymograph, or semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) were used to monitor the fluctuation of MMP-2, -8 and -28 protein and mRNA. The results showed that short-term (10min) AQE treatment (SAT) and long-term (24hr) AQE treatment (LAT) enhanced the synthesis and secretion of MMP-2, -8 and -28 protein. Phosphatidylinositol 3-kinase (PI3K) inhibitor and mitogen-activated protein kinase kinase (MEK) inhibitor were used to address the involved signal transduction pathways. We observe to that SAT and LAT are not right with leading the dependence on PI3 and MEK of function not the same of MMPs; Detected mRNA which measures MMP-2 and MMP-28 and will increase because of LAT by semi-quantitative RT-PCR method finally.
Relation:	校內外完全公開
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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