2014-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/644565摘要： 第二型似胰島素生長因子 (IGF-II) 傳訊 RNA 結合蛋白-3 (IMP-3) 乃是 IGF-II 傳訊 RNA 結合蛋白家族的一員。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。最近發現，IMP-3 在轉移的黑色素細胞癌中表現遠高於淺層黑色素細胞癌，而在良性黑色素痣則不表現，顯示 IMP-3 是一個新的黑色素細胞癌進展標記。在過去的研究中，我們發現IMP-3 表現量與黑色素細胞癌的移動、侵襲、腫瘤形成與淋巴轉移能力有關，特別是在肢端小痣型黑色素細胞癌具有更大的統計差異，顯見 IMP-3是具有本土重要性的腫瘤惡化標記。目前我們對於 IMP-3調控黑色素細胞癌的分子機轉所知有限。為了進一步釐清 IMP-3的功能和促進黑色素細胞癌進展的機轉，我們做了微陣列分析，來比較 IMP-3 高表現細胞和對照組的 mRNA 表現差異。依據微陣列分析和之前的論文，我們發現 IMP-3可能會調控一些 mRNAs，包括：HMGA2、c-MYC、TGF-β2、CCL20、WISP2、TES-85 和 TGM2，其中又以 HMGA2、TGF-β2、c-MYC 有較顯著的調控效應。因此，本計畫將探討在 IMP-3調控黑色素細胞癌進展的過程中，HMGA2所扮演的角色，以及 IMP-3調控 HMGA2與其他可能 mRNAs 表現之分子機轉，具體目標包括： 1. 運用核醣蛋白免疫沉澱技術合併次世代高通量分析來完整探討所有與IMP-3結合的RNAs； 2. 探討在黑色素細胞癌移動與侵襲過程中，IMP-3如何與HMGA2作用； 3. 確認IMP-3會影響HMGA2與其他可能受調控之RNA的穩定度與降解； 4. 確認IMP-3調控HMGA2與其他可能受調控之RNA表現之分子機轉； 5. 研究在IMP-3調控黑色素細胞癌轉移過程中，HMGA2所扮演的角色。 本研究計畫原已於去年度提出申請，因故未能獲得經費補助；我們在過去一年中增加了更多個案數，同時做了單變項與多變項分析，證實 IMP-3在亞洲人特有之肢端型黑色素細胞癌有更顯著的臨床意義；另外也完成了 RIP/qRT-PCR 實驗，確認黑色素細胞癌的 IMP-3 可與 HMGA2、TGF-β2、c-MYC的 RNA 結合，而影響其表現量。因此於今年度再次提出後續之研究計畫。藉由達成以上研究目標，我們期待能應用相關分子機轉，來建立針對亞洲人黑色素細胞癌的嶄新治療方式，因此具有其重要轉譯醫學價值。<br> Abstract: Insulin-like growth factor-II messenger RNA (mRNA)-binding protein-3 (IMP-3) is a member of insulin-like growth factor-II mRNA-binding protein family and is expressed during embryogenesis and in some malignancies. Melanoma is the most aggressive skin cancer, with a propensity to metastasize, and is resistant to most of the current therapeutic regimens. Recently, IMP-3 has been found to be a novel progression marker in malignant melanoma because it is expressed in metastatic melanomas significantly more than in thin melanomas, and is not expressed in benign nevi. Our preliminary results showed that IMP-3 levels were correlated with migration, invasion, tumorigenicity, and lymphatic metastasis ability of melanoma cells. To date, little is known about the molecular mechanisms involved in the complex regulation of IMP-3 in melanoma cells. To elucidate the function of IMP-3 and the potential mechanism of IMP-3 induced melanoma progression, we did a microarray study to compare the expression profiles of IMP-3 over-expression cells and the vector controls. According to microarray report and previous reports, we found IMP-3 could regulate several mRNAs: HMGA2, c-MYC, TGF-β2, CCL20, WISP2, TES-85 and TGM2. Among them, HMGA2, c-MYC, and TGF-β2 have been found to be highly associated with IMP-3. However, the role of HMGA2 in melanoma progression has not been well investigated. Therefore, the aim of this study is to investigate the roles of HMGA2 and other IMP-3 associated mRNAs that regulate melanoma migration, invasion, tumorigenicity, and lymphatic metastasis, as well as the molecular mechanisms of IMP-3 regulating HMGA2 expression: 1. Using RIP-deep sequencing to comprehensively study all possible RNA targets that may be interacted with IMP-3 2. To examine the interaction between IMP-3 and HMGA2 in melanoma migration and invasion. 3. To investigate mRNA stability and decay of the associated RNAs regulated by IMP-3 expression in melanoma. 4. To investigate the molecular mechanisms of IMP-3 regulating the expressions of HMGA2 and other associated RNAs. 5. To investigate the roles of HMGA2 in IMP-3 mediated melanoma metastasis. The proposal of this project was failed to be granted last year, so we increased the case number up to 141 melanocytic lesions and 97 melanomas, performing univariate and multivariate analyses to confirm that IMP-3 expression has clinical significance in acral lentiginous melanoma. We also performed RIP/qRT-PCR experiment to identify that IMP-3 is associated with RNAs of HMGA2, c-MYC, and TGF-β2 and also regulates the expression of HMGA2. The above results indicate that IMP-3 may promote melanoma progression by regulating the expression of HMGA2 or other IMP-3-associated RNAs. The long term goal of this study is to shed light on the associated molecular pathways, and to develop novel therapeutic treatment for melanoma in Asian people, especially the acral lentiginous melanoma.IMP-3侵襲黑色素細胞癌移動增生IMP-3InvasionMelanomaMigrationProliferation