2010-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/655551摘要：黑色素細胞癌是最惡性的皮膚腫瘤，它極易轉移，且對大部分的現有化學治療藥物具有抗藥性。微核醣核酸 (miRNAs) 是內生性的小片段 (≈22-nt) RNA，它可以抑制特定序列基因之表現。有越來越多的證據顯示miRNA的表現在人類癌症中出現異常，最近研究也發現miRNA與黑色素細胞癌的發生與進展有關。由於目前對於轉移的黑色素細胞癌並無有效的治療方法，找出新的分子機制或預後標記，將可以促成開發針對轉移黑色素細胞癌的新療法。在先前的初步研究中，我們選取了六位黑色素細胞癌病患的轉移部位組織與原發部位進行同一個人間的比較，運用miRNA基因微陣列分析，找到5個miRNAs (miR-211, miR-107, miR-141, miR-187 and miR-514) 在6對檢體中一致下降。其中，miR-211的相對表現量與黑色素細胞癌細胞株的移動力與侵襲力成相反關係，體外實驗也顯示miR-211可能藉由抑制黑色素細胞癌的移動與侵襲而調控其轉移。本研究將分三年探討miR-211在黑色素細胞癌轉移所扮演的角色：第一年：(1) 在病患原發腫瘤與轉移處 (共96檢體)，以及小鼠活體黑色素細胞癌轉移模式中，比較miR-211表現量之差異；(2) 運用小鼠活體黑色素細胞癌轉移模式來確認miR-211之抑制效果。第二年：(1) 以生化方式直接純化RISC與其結合的mRNA來檢測並確認miR-211之標的基因mRNAs；(2) 確認miR-211與其標的mRNAs的交互作用第三年：(1) 研究miR-211與TRPM1表現之關係；(2) 研究轉譯因子MITF調控miR-211與TRPM1表現的角色。希望藉由此研究確認miR-211在黑色素細胞癌轉移所扮演的角色與其分子機制，並可望開發未來診斷與治療的新方向。<br> Abstract: Melanoma is the most aggressive skin cancer, with a propensity to metastasize, and is resistant to most of the current therapeutic regimens. MicroRNAs (miRNAs) are endogenous ≈22-nt noncoding small RNAs, which negatively regulate gene expression in a sequence-specific manner. Increasing evidence shows that miRNA gene expression is deregulated in human cancers. Recently, several reports also found some miRNAs are involved in melanoma development. Since there is currently no effective treatment for metastatic melanoma, identifying novel molecular mechanisms and prognostic markers may lead to development of new treatments for metastatic melanomas.Our preliminary results of comparing 6 pairs of metastatic and primary melanomas from six patients revealed that 5 miRNAs (miR-211, miR-107, miR-141, miR-187 and miR-514) were down-regulated in metastatic melanomas. Further results suggested that miR-211 could regulate melanoma metastasis by inhibiting migration/invasion abilities of melanoma cells. In this project, we will investigate the roles of miR-211 in melanoma metastasis in 3 years:The 1st year:(1) Evaluate the significance and correlation of miR-211 levels with melanoma metastasis in 96 human tissues and mouse metastasis models;(2) Assess the metastasis-inhibiting properties of miR-211 in mouse models.The 2nd year:(1) Investigate and verify the down-stream targets and mechanisms of miR-211 by a novel biochemical method- combined RNA-induced silencing complex (RISC) purification with microarray analysis of bound mRNAs;(2) Confirm the binding of miR-211 to some interesting mRNA targets revealed by this biochemical method.The 3rd year:(1) Investigate the regulation of miR-211 expression and its relationship with the TRPM1 expression;(2) Investigate the roles of miR-211 in melanoma cells transfected with MITF expression vector or dominant-negative MITF expression vector.The aim of this project is to investigate the roles and molecular mechanisms of miR-211 in melanoma metastasis, which may improve the development of prognostic markers and novel treatments for melanoma.癌症轉移黑色素細胞癌微核醣核酸Cancer metastasisMelanomaMicroRNA