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研究生:黃文怡
研究生(外文):Huang Wen-Yi
論文名稱:細胞凋亡相關蛋白於口腔癌組織之表現
論文名稱(外文):Expression of apoptosis-associated proteins in oral cancer tissue
指導教授:陳振漢陳振漢引用關係
指導教授(外文):Chen Jenn-Han
學位類別:碩士
校院名稱:國防醫學院
系所名稱:牙醫科學研究所
學門:醫藥衛生學門
學類:牙醫學類
論文種類:學術論文
論文出版年:1999
畢業學年度:87
語文別:中文
論文頁數:58
中文關鍵詞:細胞凋亡口腔癌TUNEL
外文關鍵詞:apoptosisoral cancerTUNEL
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細胞凋亡是目前癌症研究的重要課題之一。其之所以重要,除了因為目前臨床上的化學及放射線治療與引發癌細胞產生凋亡有密切關係之外,如何藉研究癌細胞的凋亡機制,更可以了解癌細胞在活體中的生物特性,從而改進或研發更有效的臨床治療,是癌症研究人員的努力目標。本實驗即分析口腔癌細胞中,有關凋亡相關蛋白的表現,與癌細胞產生凋亡之間的關聯性探討。我們共取34例口腔癌病理切片,以原位TUNEL染色法偵測凋亡細胞,以免疫組織化學染色法偵查凋亡相關蛋白Fas、Fas ligand、Bax、及p53,並統計方法分析之間的相關性。結果顯示TUNEL染色法可以很靈敏地偵測到凋亡細胞,而Fas的表現是以胞質及胞膜染色為主,Fas ligand 及Bax則以胞質染色為主,而p53陽性染色是位在細胞核中。在經統計分析後,我們發現口腔癌細胞的凋亡細胞數目,(1) 隨著癌細胞的分化程度變差而下降 (p = 0.06),而其中分化良好與分化差之間有顯著地差異 (p < 0.05) ; (2) 隨著Fas之表現的增加而增多的趨勢 (p = 0.08)。另外最有顯著意義的是,癌細胞的分化程度愈差,則Fas之表現愈少(p = 0.0006)。由本實驗所獲結果可以看出,癌細胞的分化及Fas之表現,可以做為預測癌細胞凋亡的指標。而整體而言,Fas在口腔癌組織中的表現比Fas ligand少,表示口腔癌細胞可能藉由降低Fas而增加Fas ligand表現的機制,逃避由Fas/Fas ligand所引發的細胞凋亡。

Apoptosis is one of the most important topics in current cancer researches. Many reports have demonstrated that cell death of cancer cells induced by chemotherapeutic drugs and irradiation are through the process of apoptosis. Therefore, developing new treatment of cancer via apoptosis induction is the way researchers have spent much effort to achieve their goal. In this experiment, we investigate the expression of apoptosis-related proteins in oral cancer tissue and search the correlation among protein expression and the degree of apoptosis within the cancer tissues. Total 34 cases were chosen for TUNEL staining to detect the apoptotic cancer cells and for immunohistochemical staining to detect the expression of Fas, Fas ligand, Bax, and p53. The results showed that the TUNEL method could detect apoptotic cells as well as apoptotic bodies with high sensitivity. After statistics analysis among each parameter, we found that the number of apoptotic cells found in oral cancer tissue was marginally correlated with tumor histologic grade (p = 0.06) and there is significantly difference between well and poor differentiated cancer tissues (p < 0.05). The most significant result in this study was the reverse correlation between Fas expression and the tumor histologic grade (p = 0.0006). Our results suggest that the expression of Fas as well as the degree of cell differentiation may have value in assessment of tumor apoptosis. The down-regulated expression of Fas and up-regulated expression of Fas ligand of the oral cancer cells may provide a mechanism for cancer cell escaping the Fas/Fas ligand induced apoptosis in vivo.

正文目錄
前言 ----------------------------------------------------------------1
材料與方法 -------------------------------------------------------6
結果 ---------------------------------------------------------------15
討論 ---------------------------------------------------------------20
參考文獻 ---------------------------------------------------------25
表目錄
表1:中華民國八十六年臺灣地區主要癌症死亡原因--------------------------------36
表2:本實驗口腔癌之臨床資料及組織化學染色結果------------------------------- 37
表3:口腔鱗狀上皮癌的分化程度與各凋亡相關蛋白之表現情形---------------- 38
表4:口腔鱗狀上皮癌的分化程度與各凋亡相關蛋白表現之間的相關性------- 39
圖目錄
圖1:凋亡細胞在HE染色之型態---------------------------------------------------------40
圖2:正常口腔上皮與癌組織在TUNEL染色之型態---------------------------------41
圖3:凋亡小體與凋亡細胞在TUNEL染色之型態------------------------------------42
圖4:正常與分化不良的口腔上皮在Fas染色之型態---------------------------------43
圖5:Fas在角質化癌細胞中之表現-------------------------------------------------------44
圖6:正常與分化不良的口腔上皮在Fas ligand染色之型態-------------------------45
圖7:Fas ligand強弱與TIL浸潤癌組織之關係----------------------------------------46
圖8:正常與分化不良的口腔上皮在Bax染色之型態--------------------------------47
圖9:Fas在角質化與角化不全癌細胞中之表現---------------------------------------48
圖10:P53在口腔癌細胞中之表現-------------------------------------------------------49
圖11:Western blot結果分析--------------------------------------------------------------50
圖12:口腔癌細胞之凋亡細胞數目與癌細胞分化之關係---------------------------51
圖13:三種不同分化程度之口腔癌細胞中凋亡細胞數目之比較------------------51
圖14:分化良好之口腔癌細胞凋亡細胞與分裂細胞數目間之迴歸分析---------52
圖15:分化中度之口腔癌細胞凋亡細胞與分裂細胞數目間之迴歸分析---------52
圖16:分化差之口腔癌細胞凋亡細胞與分裂細胞數目間之迴歸分析------------53
圖17:口腔癌細胞凋亡細胞與分裂細胞數目間之迴歸分析------------------------53
圖18:Fas蛋白在不同分化程度口腔癌中表現情形----------------------------------54
圖19:Fas ligand蛋白在不同分化程度之口腔癌中表現情形-----------------------54
圖20:Bax之表現與癌細胞分化程度之間的關係-------------------------------------55
圖21:p53之表現與癌細胞分化程度之間的關係-------------------------------------55
圖22:口腔癌組織中癌細胞之凋亡數目與其Fas表現之關係----------------------56
圖23:口腔癌組織中癌細胞之凋亡數目與其Fas ligand表現之關係--------------56
圖24:口腔癌組織中癌細胞之凋亡數目與其Bax表現之關係----------------------57
圖25:口腔癌組織中癌細胞之凋亡數目與其p53表現之關係----------------------57
圖26:TUNEL染色法原理-----------------------------------------------------------------58

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