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研究生:王嘉鴻
研究生(外文):Chia-Huang Wang
論文名稱:MicroRNA 92a與 E-cadherin在大腸直腸癌之相關性分析
論文名稱(外文):The association between microRNA 92a and E-cadherin in colorectal cancer
指導教授:蔡崇弘蔡崇弘引用關係鄭雅文鄭雅文引用關係
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:62
相關次數:
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研究目的:大腸直腸癌全球發生率在所有癌症中排名第四位,在2008年有近61萬人死於大腸直腸癌;而微型核醣核酸(microRNAs)在後轉錄階層(post-transcriptional level)可以調控基因表現,並且在腫瘤發展、演進及轉移上扮演重要角色,本研究想了解在大腸直腸癌microRNA 92a (miR-92a) 與E-cadherin兩者相關性為何。
研究方法及資料:本研究收集了158位大腸直腸癌病患的手術切除之腫瘤組織檢體,利用即時定量反轉錄聚合酶連鎖反應 ( Real-time RT-PCR )進行miR-92a與E-cadherin基因檢測與分析。
研究結果:本研究顯示在大腸直腸癌患者中,miR-92a高表現量與淋巴結轉移具正相關(p=0.012),校正年齡、性別與分化程度下,此相關性乃具顯著性(p=0.01),而 E-cadherin表現量與淋巴結轉移並不具相關性(p=0.728),miR-92a 與E-cadherin基因兩者亦無顯著相關(p=0.056)。
結論與建議:在大腸直腸癌病患miR-92a高表現量較易發生淋巴結轉移,而E-cadherin與淋巴結轉移並不具相關性,且miR-92a與E-cadherin基因兩者亦不具相關性,miR-92a可能不是經由抑制E-cadherin基因的表現,進而促進淋巴結轉移。

Objectives: Colorectal cancer (CRC) is the fourth highest cause of cancer deaths in worldwide. On 2008, 608,700 people deaths in colorectal cancer. MicroRNAs regulate gene expression at the post-transcriptional level and play important roles in tumor development, progression, and metastasis. The aim of this study was to investigate the correlation between microRNA 92a (miR-92a) and E-cadherin expression in colorectal cancer.

Materials and methods: A total of 158 colorectal cancer patients were enrolled . MiR-92a and E-cadherin were analyzed by real-time PCR.

Results: The expression levels of miR-92a in tumor tissues were significantly positive correlated with lymph node metastasis of CRC patients (p=0.012).After adjusting for age, gender and differentiation status, this correlation remained significantly (p=0.01). In addition, expression levels of E-cadherin was not correlated with lymph node metastasis(p=0.728).The correlation of miR-92a and E-cadherin gene was not significant in this study (p=0.056).

Conclusion: We found that high expression of miR-92a was positive correlated with lymph node metastasis inCRC. But expression levels of E-cadherin was not correlated with lymph node metastasis. Down-regulation of E-cadherin gene in CRC was not associated with miR-92a up-regulation .Thus, we suggested that miR-92a involved in lymph node metastasis of CRC patients may not through E-cadherin down-regulation .

壹、 文獻綜論 1
一、 大腸直腸癌 1
1.1 流行病學 1
1.2 臨床症狀與診斷 2
1.3 癌症分期 3
1.4 疾病治療 4
1.5 疾病預後 4
二、 微型核醣核酸(microRNAs,miRNAs) 5
2.1 MicroRNAs之簡介 5
2.2 MicroRNAs之合成與作用機制 6
2.3 MicroRNAs在大腸直腸癌的表現 7
2.4 MicroRNA 92a(miR-92a)之簡介 8
2.5 MicroRNA 92a在大腸直腸癌的表現 9
三、 鈣黏著素E (E-cadherin) 10
3.1 E-cadherin之簡介 10
3.2 E-cadherin在腫瘤細胞扮演之角色 11
3.3 MiR-92a與E-cadherin相關性及其在癌症相關研究 12
貳、 研究動機 13
參、 實驗方法與材料 14
一、 化學藥品與試劑 14
二、 實驗方法 15
1.1 檢體收集 15
1.2 細胞株 16
1.3 MicroRNAs表現分析 17
1.4 標的基因表現分析 21
1.5 統計分析 24
肆、 結果 25
一、 MicroRNA 92a與大腸直腸癌臨床病理分期之相關性 25
二、 E-cadherin與大腸直腸癌臨床病理分期之相關性 26
三、 MicroRNA 92a與E-cadherin在大腸直腸癌之相關性 27
四、 MicroRNA 92a與E-cadherin在大腸直腸癌細胞株表現 28
伍、 討論 29
一、 MicroRNA 92a在大腸直腸癌的相關研究 29
二、 MicroRNAs與淋巴結轉移在大腸直腸癌之相關性 30
三、 MicroRNA 92a目標基因與生物功能 31
四、 結論 33
陸、 未來研究方向 34
柒、 參考文獻 35
捌、 圖與表 45
圖一、大腸直腸癌細胞株miR-92a與E-cadherin表現量 45
表一、大腸直腸癌病患基本資料 46
表二、MiR-92a與臨床因子之相關性 47
表三、淋巴結轉移與其他臨床因子之相關性 48
表四、淋巴結轉移與各因子之邏輯性廻歸分析 49
表五、E-cadherin與臨床因子之相關性 50
表六、MiR-92a與E-cadherin之相關性 51
表七、MiR-92a與E-cadherin相關性分析 52
玖、 附圖與附表 53
附圖一、 1979-2009年台灣大腸直腸癌發生率 53
附表一、 2010年台灣十大癌症個案數 54
附表二、 TNM staging for colorectal cancer 55
附表三、 The Dukes'' system 57
附表四、 MicroRNAs 之cDNA 合成試劑比例 58
附表五、 MicroRNAs 之cDNA 合成PCR Conditions 58
附表六、 MicroRNAs及時定量聚合酶連鎖反應試劑比例 59
附表七、 MicroRNAs及時定量聚合酶連鎖反應條件 59
附表八、 標的基因之cDNA 合成試劑比例 60
附表九、 標的基因之cDNA 合成PCR Conditions 60
附表十、 標的基因及時定量聚合酶連鎖反應試劑比例 61
附表十一、 基因Primer 序列 61
附表十二、 標的基因及時定量聚合酶連鎖反應條件 62

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