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系統識別號 U0026-1607201414202400
論文名稱(中文) 肺癌形成中Sp1調控microRNAs對CD44表現之探討
論文名稱(英文) To study the role of Sp1-mediated microRNAs in CD44 expression during lung tumorigenesis
校院名稱 成功大學
系所名稱(中) 生物資訊與訊息傳遞研究所
系所名稱(英) Insitute of Bioinformatics and Biosignal Transduction
學年度 102
學期 2
出版年 103
研究生(中文) 林新晉
研究生(英文) Shin-Jin Lin
學號 Z26014053
學位類別 碩士
語文別 中文
論文頁數 99頁
口試委員 指導教授-洪建中
口試委員-張文昌
口試委員-呂增宏
中文關鍵字 肺癌  Sp1  CD44  microRNA 
英文關鍵字 Lung cancer  Sp1  CD44  microRNA 
學科別分類
中文摘要 肺癌為世上最常見的的癌症之一,而且也是主要造成死亡的癌症。此外基因的變化(genetic alterations)及表遺傳的改變(epigenetic change)如DNA的甲基化(DNA methylation)、組織蛋白的修飾(histone modifications)及microRNA的調控等等,這些調控都參與了腫瘤的生成過程(tumorigensis)。microRNA為非編碼的單股RNA(noncoding single-stranded RNA),microRNA可以結合上mRNA 3端非轉譯區(3'untranslated regions,3'UTR s)來調控基因的表現量。我們最近研究結果顯示轉錄活化因子Sp1表現量於肺癌晚期較肺癌早期來得低,另外我們也發現在肺癌中Sp1可以藉由miR-182來負調控FOXO3,這樣的結果可以讓肺癌在晚期有更好的轉移能力。然而對於Sp1負調控肺癌細胞轉移的能力,截至目前仍尚未釐清。在此篇研究,首先我們發現了miR-182可以調控CD44,另外在靜默Sp1及miR-182的實驗結果都可以發現,CD44的蛋白質及mRNA表現量都明顯地上升,但有趣的是Sp1對於CD44的抑制效果相較於miR-182來得明顯許多,所以我們認為Sp1對於CD44的調控並非只單一藉由miR-182,而是更廣泛的緊緊調控著CD44。在本篇研究我們找到了另外兩種藉由Sp1調控進而抑制CD44的microRNA如miR-106a及miR-150。從靜默Sp1的實驗結果我們可以發現miR-106a及miR-150表現量明顯的下降,另外從我們實驗室所建立的KrasG12D條件式基因轉殖鼠(KrasG12D induced lung cancer transgenic mice model)及人類肺癌檢體,相較於肺癌組織,正常組織中CD44呈高度表現,另外Sp1則呈低度表現。實驗結果顯示在肺癌的進展過程中,Sp1與CD44呈負相關。在未來可以藉由ChIP-NGS來發掘出更多Sp1所能調控的microRNA,此外也會去探討CD44不同的變異型在癌症進展過程所扮演的角色。綜合以上結論,在肺癌進展過程中,Sp1可以藉由miR-106a、miR-150及miR-182進而負調控CD44的表現量。此研究也提供了Sp1在癌症進展過程中對於標的基因調控的新視野。

英文摘要 We showed that miR-106a and miR-15 are Sp1- activated miRNAs, whose expression increased in lung cancer. On the other hand, in the early stages of lung cancer progression, Sp1 stimulates miR-106a and miR-150 expression, which in turn decreases CD44 expression. This stimulates proliferation and tumor growth. In the late stages, Sp1 and miR-106a and miR-150 decline, thus increasing CD44 expression, which leads to lung metastasis. This study provides a new vision for Sp1-mediated target gene expression to modulate cancer progression.
Key words: Lung cancer; Sp1; CD44; microRNA
論文目次 目錄
中文摘要 ………………………………………………………I
英文摘要 ………………………………………………………II
縮寫檢索表 ……………………………………………………V
致謝 ……………………………………………………………VI
目錄 ……………………………………………………………VII
第一章 序論 ………………………………………………… 1
第一節 前言 ……………………………………………………1
第二節 肺癌 ……………………………………………………1
第三節 Specificity protein 1 (Sp1) ……………………………6
第四節 microRNA ……………………………………………8
第五節 Cluster differentiation 44 (CD44) ……………………10
第二章 研究動機與目的 ……………………………………13
第三章 實驗材料 ……………………………………………14
第四章 實驗方法 ……………………………………………22
第五章 實驗結果 ……………………………………………40
第六章 實驗討論 ……………………………………………51
第七章 參考文獻 ……………………………………………56
第八章 附圖 …………………………………………………62
第九章 附錄 …………………………………………………88
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