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研究生(外文):Chen-Hsin Chen
論文名稱(外文):The Role of miR-20a in Human Prostate Cancer Cell Death by DNA-topoisomerase Inhibitors
指導教授(外文):Yat-Pang Chau
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微核醣核酸(microRNAs or miRNA)是一個長約20~23核苷酸的小片段之內生性RNA,miRNA的特殊排列順序可與標靶基因 3'UTR結合,通過降解或抑制mRNA的翻譯,藉此達到調控基因表現的目的和抑制蛋白質的生產,在基因中的調控上扮演重要的角色。最近文獻指出,在人類的基因中有超過三分之一被miRNA所調控,其中在癌症的轉移或是腫瘤的生長研究中發現某些miRNA出現突變或不正常的表現,推測這些miRNA可能扮演著抑癌或是致癌的功能。有鑑於前列腺癌個案在台灣漸趨增加,因此預防與治療前列腺癌在醫學研究上十分重要。已知DNA拓樸異構酶的抑制物(DNA topoisomerases inhibitors)可以干擾細胞的DNA合成,臨床上DNA拓樸異構酶的抑制物如Doxorubicin及Camptothecin皆被廣泛用來當做抗癌藥物使用,因此我們有理由的推測這些藥物會影響細胞內miRNA的表現。在本篇論文中,我們主要針對DNA拓樸異構酶的抑制物對前列腺癌細胞內miRNA的表現影響和差異,並且深入探討miR-20a在此條件下所扮演的角色。
我們發現Camptothecin處理後的前列腺癌細胞的miR-20a表現量比Doxorubicin處理後有非常明顯的上升,miR-20a的下游標的基因E2F-1和cyclin E1也隨著下降,加入anti-miR-20a於前列腺癌細胞中,抑制miR-20a的表現會降低前列腺癌細胞經處理抗癌藥物Doxorubicin及Camptothecin後的細胞存活率。這些結果顯示miR-20a表現抑制後,前列腺癌細胞DU145和PC3對DNA拓樸異構酶抑制物的藥物敏感性上升、耐受性下降,抗癌藥物在較短作用時間內可以達到較高致死效率,增加藥物的抗癌效果。因此我們認為miR-20a在前列腺癌細胞的基因調控機制和對抗癌藥物的敏感性等方面扮演著重要的角色。
MicroRNAs (miRNAs) are endogenous RNAs of 20-23 nucleotides in length that negatively regulate the expression of genes by binding to the 3’-untranslated region (UTR) of the target mRNAs. Recent studies suggest that up to one third of human genes are regulated by miRNAs. MicroRNAs may act as either tumor suppressor or oncogenic factor in the process of carcinogenesis and tumor growth. Because of the increase of prostate cancer incidence in Taiwan, prevention and treatment of prostate cancer is an important event for biomedical secientists. DNA-topoisomerase inhibitors such as doxorubicin (DOX) and�n camptothecin (CPT) that cause DNA damage have been widely used as anticancer drugs. It is reasonable to surmise that these drugs may be able to alter the expression of miRNA genes. In this study, we aim to examine the change of miRNAs profile and the role of miR-20a in human prostate cancer cell lines following the treatment without or with DNA topoisomerase inhibitors.
We here found the increased level of miR-20a expressions in DU145 and PC3 cells after CPT treatment compared to doxorubicin treatment, respectively. Therefore, we examined the role of miR-20a in the mechanism of CPT- or DOX-induced prostate cancer cell death. To elucidate the role of miR-20a, downregulation of miR-20a was tested in DU145 and PC3 cells. Our data showed that suppression of miR-20a with anti-miR20a significantly decreased the survival rate of DU145 and PC3 after exposure to DOX and CPT. The results suggested that downregulation of miR-20a may increase the susceptibility and decrease the drug resistance of prostate cancer cells to topoisomerase inhibitors. Therefore, we conclude that miR-20a may play a crucial role in the gene regulation and the drug susceptivity of prostate cancer cells.
中文摘要 i
Abstract ii
目錄 iii
第一章 緒論(Introduction) 1
一、MicroRNA (miRNA) 1
二、前列腺與前列腺癌 7
三、抗癌藥物概述 13
第二章 目的(Purpose) 17
第三章 實驗材料與方法(Material and Method) 18
第四章 結果(Result) 26
第五章 討論(Discussion) 33
第六章 結論(Conclusions) 38
第七章 參考文獻(Reference) 39
結 果 51
附 錄 80
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