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研究生:柯順耀
研究生(外文):Shun-Yao Ko
論文名稱:類澱粉前趨蛋白質之相關研究:I.神經細胞中檳榔子水萃取物對於類澱粉前趨蛋白質修飾之調控II.類澱粉前趨蛋白質在口腔鱗狀上皮細胞癌中扮演的角色
論文名稱(外文):The Research of Amyloid Precursor Protein: I. Amyloid Precursor Protein Processing Is Regulated by Areca Nut Extract in SK-N-SH Cells II. The Role of Amyloid Precursor Protein in Oral Squamous Cell Carcinoma
指導教授:劉宗榮劉宗榮引用關係
指導教授(外文):Tsung-Yun Liu
學位類別:博士
校院名稱:國立陽明大學
系所名稱:藥理學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:167
中文關鍵詞:ANEAPP阿茲海默症OSCC綠茶ADAM 10
外文關鍵詞:ANEAPPAlzheimer’s diseaseOSCCGreen teaADAM 10
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本論文主要探討檳榔子水萃取物(ANE)對神經瘤細胞株SK-N-SH中類澱粉前趨蛋白質(APP)之調控及探討APP在口腔鱗狀上皮細胞癌(OSCC)中扮演的角色。SK-N-SH細胞分別處理ANE、濃縮分離後之ANE (fANE)和arecoline,觀察APP mRNA、蛋白質和APP修飾後產物sAPP的表現。結果發現ANE和fANE可減少sAPP,arecoline可增加sAPP。動物實驗使用天竺鼠餵食ANE,也發現在腦脊液中sAPP的表現會被抑制。這些結果表示ANE可能具有治療阿茲海默症的潛力。APP已被證實可以調控細胞增殖,以OSCC和配對之癌旁正常組織(NCMT)萃取RNA和蛋白質,觀察APP表現和OSCC的關係。RT-PCR分析結果顯示APP mRNA的表現量在OSCC中較NCMT有顯著增加,Immunohistochemistry (IHC)分析結果也和RT-PCR結果相同。比對病患的臨床資料發現,APP mRNA表現增加之OSCC病患存活率顯著的降低。使用口腔癌細胞株OEC-M1處理APP反意核酸抑制細胞中APP的表現,結果發現APP反意核酸抑制APP的表現,進而抑制細胞生長。在MBN處理中國倉鼠之口腔癌動物模式中,APP大量表現於OSCC組織。使用綠茶萃取物及綠茶主要的成份EGCG處理OEC-M1和MBN誘發中國倉鼠口腔癌,結果發現EGCG可以抑制OEC-M1細胞中APP mRNA、蛋白質和sAPP的表現。另外,綠茶萃取物可以顯著降低MBN誘發的口腔癌中APP的表現。這些結果意味著APP對於口腔癌化扮演一個重要的角色。
最近的研究報告指出具有α-secretase活性修飾APP之ADAM 10在許多的腫瘤中皆有發現。因此,ADAM 10在OSCC中可能和APP具有相關性。IHC分析結果發現ADAM 10的表現在OSCC中較NCMT有顯著增加,RT-PCR結果也和IHC相同。另外,統計的結果發現,在OSCC中APP及ADAM 10兩個基因的表現有正相關性且具有顯著意義。OEC-M1處理ADAM 10反意核酸後抑制ADAM 10的表現,同時也抑制細胞的生長。但是ADAM 10不影響sAPP的表現,此結果表示ADAM 10在OEC-M1中可能不具α-secretase活性。以上結果暗喻著ADAM 10在口腔癌化中也相當的重要。
This study is designed to test the effects of areca nut extract (ANE) on the expression of amyloid precursor protein (APP) in SK-N-SH cells, and the role of APP in OSCC. SK-N-SH cells were treated with ANE, fractionated ANE (fANE) and arecoline. The results demonstrated that ANE and fANE decreased sAPP in medium. On the other hand, arecoline increased the sAPP expression. The in vivo treatment of guinea pig with ANE also decreased sAPP in cerebrospinal fluid. These results suggest that ANE might have potential in the development of drugs for Alzheimer’s disease. APP has been demonstrated to regulate cell proliferation. This study also tested the relationship between APP expression and the development of oral squamous cell carcinoma (OSCC). RT-PCR analysis showed increase of APP mRNA in OSCC as compared to the non-cancerous matched tissue (NCMT). Immunohistochemistry analysis further showed a remarkable increase of APP in OSCC tissue relative to NCMT. The results also revealed that OSCC patients exhibiting increase in APP mRNA expression had significantly lower survival rate compared to patients exhibiting the opposite status. Treatment of OEC-M1 oral cancer cells with antisense oligonucleotide against APP resulted in inhibited cell proliferation as well as APP expression. The expression of APP was also highly induced in MBN-induced oral cancer in hamster buccal pouch. This study also tested the effect of green tea extract and EGCG, an important green tea polyphenol, in OEC-M1 cells and in MBN-induced hamster oral cancer. The results demonstrated that EGCG reduced the expression of APP by RT-PCR and Western blot in OEC-M1 cells. In addition, green tea extract significantly reduced the APP expression in MBN-induced oral cancer by immunohistochemistry. The above results suggest that APP might play an important role in OSCC carcinogenesis.
Recent studies have revealed the expression of ADAM 10 in many tumors. ADAM 10 has been known to have α-secretase activity, and therefore might be involved in APP processing. The relationship between the expression of APP and ADAM 10 was also tested in this study. Immunohistochemistry demonstrated that ADAM 10 was highly expressed in OSCC. RT-PCR analysis revealed that the expression of ADAM 10 in OSCC was higher than that in NCMT. In addition, the expression of ADAM 10 was correlated to the expression of APP in OSCC. In vitro treatment of antisense oligonucleotide against ADAM-10 in OEC-M1 cells inhibited cell growth. However, the sAPP expression was not inhibited by ADAM 10 antisense oligonucleotide. This suggests that ADAM 10 might not have α-secretase activity in OEC-M1 cells. Nevertheless, ADAM 10 might still be involved in OSCC carcinogenesis.
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