臺灣博碩士論文加值系統

細胞分化的過程牽涉到一群特定的基因有計畫的表現，為了找出參與表皮分化的基因，我們使用了互補去氧核糖核酸微陣列分析(cDNA microarray)的技術，來分析角化及未角化的C9細胞基因表現的差異，並且以反轉錄聚合酶鏈鎖反應(RT-PCR)的技術再次確定其結果。微陣列分析的結果顯示，C9細胞株分化的模式符合已知的表皮分化模式，因此，C9細胞株是一個適合拿來研究表皮分化的模式(model)。我們也發現了一些可能與表皮分化相關的轉錄因子，諸如TLE1、KLF4、CSRP和CREG等基因，這些基因皆表現於表皮或神經組織，大部分是轉錄抑制者。表皮和神經之所以會有共同的轉錄調節因子，很可能是起因於他們皆來自胚胎發育時的外胚層，這也許是研究表皮分化的一個新方向。未來，我們可以用免疫螢光染色或組織原位雜交等技術來確定這些基因表現的位置，以直接證明這些基因與表皮分化的關係。

Cellular differentiation is a process in which a group of specific gene is programmatically induced. In order to find out genes that participate in epithelium differentiation, we used cDNA microarray technique to analyze the difference of gene expression between keratinized and non- keratinized C9 cells, and recheck the data by RT-PCR. The microarrray data show that the pattern of gene expression in C9 cell differentiation is consistent with previously studies of epidermal differentiation. Hence, C9 cell is a potential model for studying epidermal differentiation. We also find some interesting transcription factors related to cell differentiation, such as TLE1, KLF4, CSRP and CREG gene. Most of those genes, expressing in epidermis or neurons, are transcription repressor. The epidermis and neurons have the same transcription factors probably because both of them come from the ectoderm during embryo development. It’s a new direction to research epidermis differentiation. In the future, we can try to locate the expression sites of those genes by immuno-fluorescence staining or in situ hybridization to prove the relationship between those genes and epidermis differentiation directly.

目錄………………………………………………………….……………I
圖表目錄………………………………………………………………..III
附錄目錄………………………………………………………………..IV
中文摘要………………………………………………………………...V
英文摘要………………………………………………………………..VI
本文
壹、緒論…………………………………………………………………1
一、表皮分化……..……………………………………………………1
二、微陣列分析………………………………………………………..4
貳、材料與方法………………………………………………………….7
一、實驗材料…………………………………………………………..7
1. 腫瘤細胞株……………………………………………………..7
2. 裸鼠……………………………………………………………..7
3. 實驗藥品與試劑………………………………………………..7
4. 重要儀器………………………………………………………..8
二、實驗方法…………………………………………………….…….9
1. 腫瘤細胞解凍…………………………………………………..9
2. 腫瘤細胞繼代培養……………………………………..……..10
3. 黴漿菌測試……………………………….…………….……..10
4. 裸鼠皮下注射…………………………………………………10
5. 石蠟包埋及H&E染色……………………………..…………11
6. cDNA微陣列分析…………………………………….………11
7. clone檢定……………………………………………...………17
8. 反轉錄 聚合酶鏈鎖反應…………………….……………….17
叁、結果………………………………………………………………..22
一、細胞培養與黴漿菌測試……………………...………………….22
二、腫瘤組織切片…………………………………...……………….22
三、基因微陣列分析…………………………………...…………….23
四、clone確定…………………………………………...……………24
五、反轉錄 聚合酶鏈鎖反應……………...……………...…………24
肆、討論………………………………………...……………………...26
伍、圖表………………………………………………………………..32
陸、參考文獻……………………………………………...…………...48
柒、附錄………………………………………………………………..52

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