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研究生:張明智
研究生(外文):MING-CHIH CHANG
論文名稱:口腔鱗狀上皮細胞癌患者在周邊血液單核細胞之細胞激素mRNA表現之研究
論文名稱(外文):The mRNA expression of cytokine in cultured peripheral blood mononuclear cells from patients with squamous cell carcinoma.
指導教授:謝天渝謝天渝引用關係張基隆張基隆引用關係
指導教授(外文):TIEN-YU SHIEHKEE-LUNG CHANG
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:牙醫學研究所
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:2001
畢業學年度:89
語文別:中文
論文頁數:46
中文關鍵詞:口腔鱗狀上皮細胞癌細胞激素單核細胞腫瘤壞死因子轉型生長因子干擾素
外文關鍵詞:SQUAMOUS CELL CARCINOMACYTOKINEMONONUCLEAR CELLTUMOR NECROSIS FACTORTRANSFORMING GROWTH FACTORINTERFERON
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目前口腔癌已列入國人十大癌症死因之一,以男性而言,高居第五位。對個人、社會、乃至於國家著實造成極大負擔。嚼食檳榔與口腔癌罹患率之相關性已獲證實。因此,嚼食檳榔之口腔癌患者其免疫功能變化情形,深具研究意義。目前多數研究均偏重探討口腔癌患者體內細胞激素之變化,而鮮少著墨於細胞激素mRNA之表現調控。本論文主要利用反轉錄聚合酶鍊鎖反應 ( reverse transcription -polymerase chain reaction , RT-PCR ) 來探討具嚼食檳榔習慣之口腔鱗狀細胞癌患者周邊血液單核細胞內細胞激素mRNA 之表現,包括TNF-a、TGF-b 及IFN-g。
實驗設計上,將正常人及具嚼食檳榔習慣之口腔鱗狀細胞癌患者血液中之單核細胞加以分離,並於37oC、5% CO2 條件下,培養72 小時。經離心收集下層細胞,萃取純化 RNA,進行反轉錄 ( reverse transcription ) ,續加入 TNF-a、TGF-b或IFN-g 之特異性引子 ( primer ),於適當溫度下,進行35回合之PCR。將產物進行洋菜膠電泳( agarose gel electrophoresis ),利用溴化乙錠 ( ethidium bromide ) 染色,於紫外線下進行觀察並照相記錄。實驗結果顯示,具嚼食檳榔習慣並罹患口腔癌之患者,其周邊血液單核細胞內TNF-a 及TGF-b之 mRNA表現較正常人於統計上有意義地上升;相反的, IFN-g之mRNA表現則較正常人於統計上有意義地下降。由本論文之結果可推測,具嚼食檳榔習慣並罹患口腔癌之患者,其與免疫功能相關之細胞激素TNF-a、TGF-b 及IFN-g 之表現變化,除蛋白質層次( protein level )外,在mRNA 層次亦有相當的貢獻。至於其調控機能為何,值得進一步深入探討。
According to a 1999 report announced by the Ministry of Health, the mortality of male oral cancer patients was one of the top 10 with the highest annual increase rate, 9.5%, in Taiwan. It has been demonstrated that there is a positive rationale between betel quid chewing and the incidence of oral cancer development. Therefore, it is pivotal to investigate the variation of immune function in oral cancer patients with betel quid chewing habit. Furthermore, most of the scientific reports were focused on the profiles of cytokine protein expression in sera from oral cancer patients. So far, the regulation of cytokine mRNA expression was not clearly discovered. In this study, using reverse transcription - polymerase chain reaction ( RT-PCR ), we carried out series investigations for the cytokine mRNA expression, such as TNF-a TGF-b and IFN-g in peripheral mononuclear cell purified from oral scuamous cell carcinoma ( SCC ) patients with betel quid chewing habit.
Mononuclear cell purified from normal or betel quid users with SCC were cultured in 37oC humidified incubator with 5% CO2 for 72 h. The RNA was extracted and subjected for RT-PCR with TNF-a TGF-b or IFN-gspecific primer set. After 35 cycles amplification in an optimal temperature, the products of RT-PCR were subsequently analysed by agarose gel electrophoresis and photographed under UV-light with ethidium bromide ( EtBr ) staining. The results revealed that mRNA expression of TNF-a and TGF-b were elevated significantly in peripheral mononuclear cell from betel quid users with SCC. However, mRNA expression of IFN-gwas vice versa. Based on our results, the expression regulation of immune function related cytokines, such as TNF-a TGF-b and IFN-gwere not only in protein level, but also in mRNA level in these SCC patients. Nevertheless, the mechanism of gene regulation is worthy for further investigation.
壹、摘要------------ 1
中文摘要---------- 1
英文摘要---------- 3
貳、緒論------------ 5
參、文獻回顧---------- 8
肆、研究方法---------- 14
伍、結果------------ 22
陸、討論------------ 28
柒、參考文獻---------- 34
捌、圖表------------ 41
柒.參考文獻
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