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系統識別號 U0026-2408201211154400
論文名稱(中文) MicroRNA-193a-5p透過ERBB2以抑制鱗狀上皮細胞癌的生長及促進放射線治療的敏感度
論文名稱(英文) MicroRNA-193a-5p suppresses tumor proliferation and enhances radio-sensitivity through ERBB2 in esophageal squamous cell carcinoma
校院名稱 成功大學
系所名稱(中) 臨床醫學研究所
系所名稱(英) Institute of Clinical Medicine
學年度 100
學期 2
出版年 101
研究生(中文) 葉清彤
研究生(英文) Ching-Tung Yeh
學號 s96994015
學位類別 碩士
語文別 英文
論文頁數 78頁
口試委員 指導教授-呂佩融
口試委員-王憶卿
口試委員-劉校生
中文關鍵字 miR-193a-5p  食道鱗狀上皮癌  放射線治療 
英文關鍵字 miR-193a-5p  esophageal squamous cell carcinoma  radiotherapy 
學科別分類
中文摘要 近年來因早期診斷或治療的開發使許多癌症的存活率提高,但食道癌的五年存活率僅只有20%。許多研究指出微小核糖核酸的表現異常與癌症的形成有關,而微小核糖核酸會結合目標信使核糖核酸的3端非轉譯區,進而造成標的基因的裂解,而達到抑制該蛋白表現量的後轉譯表現。在本研究結果中,我們假設microRNA的功能異常會導致食道癌的進程,因此期望找到一群具有抑制癌細胞生長或轉移的microRNAs,首先利用微小核糖核酸晶片來了解,在食道癌組織與正常上皮組織中microRNAs表現量的差異性,結果顯示有許多微小核糖核酸的表現異常與癌症形成有關,因此依據目前已發表的文獻數目及其可能調控的下游基因,進而挑選出4個可能的抑癌微小核糖核酸作為進一步的探討,分別為miR-193、miR-200b、miR-200c、miR-451,並利用即時聚合酶連鎖反應以印證晶片結果,上述兩個結果皆顯示,這四個microRNAs在癌症組織中有低度的表現量,但只有miR-193a-5p具有成為臨床預後分析的能力,因此進一步了解miR-193a-5p在食道癌中的角色,先利用原位雜交來偵測 miR-193a-5p在組織病理切片中的表現量,結果顯示miR-193a-5p在75%的病人中有的低度的表現量,並且miR-193a-5的表現量與癌症進程有著逆相關性,所以利用增加癌細胞株中 miR-193a-5p 的表現量後,觀察miR-193a-5p在食道癌中所扮演的角色,結果現是miR-193a-5p 會直接地與ERBB2的3端非轉譯區進行結合,進而抑制ERBB2的表現量,並且透過影響ERBB2的表現量,使得癌細胞的生長速率、低生長因子、懸浮環境之生長能力下降,以及增加對於放射線治療感受性能力,而且在臨床檢體也顯示ERBB2與miR-193a-5p的表現量有著逆相關性,但是回復ERBB2的表現量後,能抑制miR-193a-5p所導致的癌細胞生長的下降,與由以上結果顯示 miR-193a-5p 具有抑制癌症進程的能力
英文摘要 Esophageal cancer is the ninth leading cause of cancer death in Taiwan. Esophageal squamous cell carcinoma (ESCC) is usually managed with combined modalities, but the prognosis for ESCC patients is still dismal. MicroRNAs (miRNAs) have been implicated in a variety of human cancers progression, and their expression signatures can provide insight into the diagnosis and prognosis of human cancers. miRNAs are small noncoding RNA molecules that bind to complementary sequences in the 3’ untranslated region (UTR) or internal region of target messenger RNAs (mRNAs) and regulate gene expression by the cleavage of target mRNAs and/or translational inhibition. In this study, we hypothesize dysregulated microRNAs promoted tumor progression. First, we analyzed the expressions of 1097 miRNAs (717 human and 380 novel miRNAs) in nine pairs of ESCC and the corresponding normal parts with NCodeTM Human miRNA Microarray V3. Second, according to the numbers of published papers and potential oncogenic targets, we selected four candidate tumor suppressive microRNAs (miR-193a-5p, miR-200b, miR-200c, and miR-451). We first examined their expressions in twenty eight ESCC and corresponding normal tissues with quantitative real time polymerase chain reaction. These four tumor suppressive microRNAs were down-regulated in tumor tissue. Because low expression of miR-193a-5p but not the others (miR-200b, miR-200c, and miR-451) in ESCC was correlated with poor prognosis of ESCC patients, we further examined the expressions of miR-193a-5p in paraffin-embedded ESCC and the corresponding normal tissues with in situ hybridization. MicroRNA-193a-5p was down-regulated in esophageal tumor of around 75% patients when compared with the normal parts. There was an inverse correlation between miR-193a-5p and tumor stage. In addition, low miR-193a-5p expression was associated with poor prognosis in 94 ESCC patients. These data indicated that miR-193a-5p played an important tumor suppressive role in ESCC. Thus, we overexpressed miR-193a-5p in ESCC cancer cell lines for detecting the tumor suppressive role of miR-193a-5p. We observed miR-193a-5p increased radio-sensitivity and decreased cell proliferation in ESCC cell lines. Tumor growth was reduced by miR-193a-5p in mouse tumor model. Next, we showed miR-193a-5p directly targeted 3’UTR of ERBB2 and repressed ERBB2 expression in vitro. An inverse correlation between ERBB2 miR-193a-5p expressions was demonstrated in ESCC cell lines and in clinical specimens. ERBB2 expression was also positively associated with tumor stage. Restored the ERBB2 expression abolished the effects of miR-193a-5p on cell proliferation and radio-sensitivity. These results indicated that miR-193a-5p played a tumor suppressive role through suppressing ERBB2, expression in esophageal squamous cell carcinoma.
論文目次 Abstract …………………………………… i
中文摘要 ……………………………… iii
Table of contents …………………… vi
Abbreviations ………………………… vii
Introduction …………………………………… 1
Materials and Methods ………………………… 10
Results …………………………………………… 22
Discussion and conclusion ………………… 30
References ………………………………… 35
Tables and figures …………………… 43
Appendix ……………………………… 74
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