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研究生:張嘉羽
研究生(外文):Chia-Yu Chang
論文名稱:胞外體CD98在肺癌細胞中的生物功能
論文名稱(外文):The biological function of exosomal CD98 in lung cancer cells
指導教授:陳逸夫陳逸夫引用關係
指導教授(外文):Yi-Fu Chen
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:生物科技學系碩士班
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2020
畢業學年度:108
語文別:中文
論文頁數:73
中文關鍵詞:肺癌肺癌細胞NL20胞外體CD98
外文關鍵詞:lung cancer cellslung cancerNL20exosomesCD98
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肺癌是全球和台灣常見的癌症之一,其五年存活率是所有癌症中最低的。近年來,由於抽菸以及空汙問題日益嚴重造成肺癌的罹癌率逐年升高,這已經成為一個迫切需要解決的問題。胞外體是一種由細胞分泌的液泡,其內包裹著蛋白質、mRNA、microRNA以及dsDNA來影響周遭細胞以達到細胞間的訊息溝通,在近年來的研究中指出:胞外體在癌症系統中扮演著重要角色。因此,我們假設肺癌細胞(H1299)的胞外體的內容物與正常肺部細胞(NL20)的胞外體不同,而其中的蛋白質差異是否反應細胞的癌化現象值得探討。首先,我們利用超高速離心機分離出兩種細胞的胞外體,並且利用質譜儀分析存在於兩者之蛋白質,結果顯示蛋白質CD98在H1299的胞外體中含量高於NL20,後續利用西方墨點法確認後也得到相同的結果。而CD98在先前的研究中已經被證實會促進細胞增生及遷移能力。因此我們將H1299的胞外體處理NL20細胞,並且根據結果發現H1299胞外體會促進NL20細胞增生及遷移,證實了先前的研究,但是效果並不明顯。為此我們利用基因選殖技術將CD98接到表現載體上,再將質體送入細胞中使其大量表現,接著收集分離其胞外體以處理NL20細胞,並發現大量表現CD98的胞外體會更加促進NL20的增生及遷移,並且是透過活化Akt及FAK從而促進此細胞生理現象。 
Lung cancer is the most common cancer worldwide and in Taiwan, and the survival rate is almost the lowest of all cancers. In recent years, because of smoking and air pollution problems, lung cancer prevalence increases year by year, which has become an urgent issue to care about. Recently, more and more studies show that exosomes play an important role in intercellular communication in cancer. Exosomes are small (50~200 nm) extracellular vesicles packed with protein, mRNA and dsDNA to affect other cells. In this study, we hypothesized that the exosomes of lung cancer cells (H1299) contain different protein contents from normal bronchial epithelial cells (NL20). First, we isolated the exosomes of the two cells, and analyzed the difference in protein content between them by mass spectrometer. We found that the score of CD98 in the exosomes of H1299 was significantly higher than NL20, and the expression level was further confirmed by Western blotting. Since CD98 has been shown in previous studies to promote cell proliferation and migration, we therefore investigated whether H1299 exosomes could confer such biological effects. The results showed that H1299 exosomes could promote the proliferation and migration of NL20 cells; but the effects were not significant. Thus, we cloned the CD98 gene to the expression vector, and overexpressed it in H1299 cells, and then isolated their exosomes to treat NL20 cells, and found that CD98-loaded exosomes promote the proliferation and migration of NL20 presumably through the activation of Akt and FAK.
目錄
摘要 1
Abstract 2
致謝詞 3
Introduction 前言 5
Material & Methods實驗材料及方法 16
Results 結果 39
Conclusion & Discussion 結果與討論 55
References 參考資料 59
Supplementary Information補充資料 67
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