論文名稱:

以二維電泳分析經檳榔萃取物處理後口腔上皮細胞之蛋白質變異

 

Identification of proteomic alterations in oral epithelial cells associated with areca asing two-dimensional electrophoresis

研究生:

林芳曲  Fang-Chu Lin

 

(以作者名查詢陽明大學館藏系統)

 

(以作者名查詢全國圖書書目資訊網)

指導教授:

張國威  Kuo-Wei Chang

        學位類別:

碩士

        學校名稱:

國立陽明大學

系所名稱:

口腔生物研究所

            學號:

38917003

          學年度:

90

          語文別:

中文

          出版年:

91

關鍵字:

蛋白體學  proteomic

 

二維電泳  two-dimensional electrophoresis

 

口腔上皮細胞  oral epithelial cell

 

檳榔萃取物  areca nut extract

全文說明:

電子全文

        論文頁數:

71

摘要:

台灣及許多東南亞國家中檳榔的嚼食習慣與口腔癌的發生有極高的相關性,然而兩者間的分子生物學證據仍未被掌握。在先前的研究中雖已發現檳榔對細胞及組織的傷害和誘導某些蛋白質表現量上的差異,但並沒有以高產能的方式探討其對細胞中蛋白質表現的直接影響。本研究使用蛋白體學研究方法――二維電泳搭配質譜儀分析法,探討口腔細胞受檳榔萃取物影響之蛋白質表現的改變,期望藉由高產能的策略探討檳榔與口腔癌間的相關性。 我們以口腔鱗狀上皮細胞癌細胞株OECM-1作二維電泳分析,並以質譜儀分析鑑定出數個表現量高之蛋白質,包括Glutathione s-transferaseProhibitinThioredoxin peroxidase 1Protein disulfide isomerase ER60及熱休克蛋白27。在以10 μg/ml濃度之檳榔水萃取物處理18小時後,可於二維電泳結果中觀察到許多蛋白質表現量上的明顯差異(大於50 %的變量),但由於這些蛋白質的表現量均偏低,在以質譜儀分析時有一定程度的限制,而在蛋白質種類鑑定上仍顯困難,因此尚未比對出確切的相對蛋白質。藉由改善蛋白質處理及質譜儀相關步驟以確定差異性蛋白質之種類,期找出表現受檳榔調控之蛋白質,並希望能建立起OECM-1的二維電泳圖譜以提供今後口腔癌相關之研究資訊,進而更確立檳榔與口腔癌化之關係。

   
 

The high correlation between areca chewing and oral squamous cell carcinomaOSCCin Taiwan and many other South Asian country was known. However the direct evidence of areca associated carcinogenesis has not been classified. Some previous experiments have shown that areca ingredients can cause damages of tissue and oral cells and the changes in expression of some proteins. But there is no research using high throughput strategies to establish the proteomic data. We use proteomic approachtwo-dimensional electrophoresis and mass spectrometry to detect the proteins expression changes modulated by areca nut extractANEtreatment and attempted to link the correlation between OSCC and areca use. We analyzed total proteins of an OSCC cell lineOECM-1 and have defined that five proteins, Glutathione s-transferaseProhibitinThioredoxin peroxidase 1Protein disulfide isomerase ER60 and heat shock protein 27 exhibited high expression in OECM-1. After ANE treatment for 18 h at 10μg/ml, we identified remarkable changes±50%on electrophoretic spots. Since these spots were low copy number proteins, there have been limits in our mass spectrometry analysis and database searching, causing difficulties in protein definition. In future our efforts will be imposed to improve mass spectrometry related steps and the maintenance of protein intensity to find out the direct correlation between protein expression and areca use. We also hope to establish 2D pattern of OECM-1 to further resolute the relationship between areca use and oral carcinogenesis.