2013-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656682摘要：惡性黑色素瘤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。SLURP-1為尼古丁乙醯膽鹼受體(nAchR)之促效佐劑並且對於表皮分化有重要影響。我們首位發現具有SLURP-1（G86R）基因突變之Meleda家族成員除了角化異常外，T細胞之活化能力受損，且無法釋出正常的SLURP-1蛋白；加入正常的SLURP-1蛋白，可使T細胞之活化能力恢復正常。四位Meleda病患中有兩位發生惡性黑色素瘤，我們也初步發現抽取三位Meleda病患的週邊單核球mRNA都可偵測到黑色素瘤幹細胞標記。而過去的文獻中顯示黑色素瘤幹細胞於惡性惡性黑色素瘤的形成及轉移扮演重要角色。因此我們假設正常的SLURP-1蛋白可經由抑制黑色素瘤幹細胞來抑制惡性惡性黑色素瘤的形成及轉移。由於目前對於轉移的惡性黑色素瘤並無有效的治療方法，本研究的目的在於找出新的分子機制，以促成開發針對轉移惡性黑色素瘤的新療法。本研究將分三年探討SLURP-1基因在惡性黑色素瘤進展及惡性黑色素瘤幹細胞所扮演的角色：第一年：(1) 探討不同時期的惡性黑色素瘤組織檢體中其SLURP-1表現與黑色素瘤幹細胞標記、及乙醯膽鹼受體(nAchR)之表現，與臨床分期及預後之關係。(2) 比較SLURP-1在有黑色素瘤幹細胞標記（如ABCB5+）, 與無黑色素瘤幹細胞標記（如ABCB5-）之 melanoma cells之表現。(3) 分析於不同惡性黑色素瘤細胞株之anti-apoptosis, angiogenesis, migration, invasion, melanoma stem cellss formation 與正常SLURP-1蛋白表現量之關聯。第二年：(1) 分析惡性黑色素瘤細胞/黑色素瘤幹細胞對正常T細胞之活化及SLURP-1表現之影響。(2) 確認抑制SLURP-1對於免疫不全小鼠活體內異體移植之惡性黑色素瘤細胞之腫瘤形成、血管生成、轉移模式及黑色素瘤幹細胞生成等生物特性之影響。第三年：(1) 研究抑制SLURP-1蛋白之表現時，對於惡性黑色素瘤細胞株生物特性之影響 (腫瘤生成, 抗細胞凋亡, 血管生成, 黑色素瘤幹細胞生成)(2) 研究增加SLURP-1蛋白表現對於惡性黑色素瘤進展之抑制的角色。希望藉由此研究確認SLURP-1在抑制惡性黑色素瘤生成、轉移及黑色素瘤幹細胞，所扮演的角色與其分子機制，並可望開發未來治療的新方向。<br> Abstract: Melanoma is the most aggressive skin cancer, with a propensity to metastasize, and is resistant to most of the current therapeutic regimens. SLURP-1 (lymphocyte antigen 6/urokinase-type plasminogen activator receptor related protein-1) is an allosteric agonist to the nicotinic acetylcholine receptor (nAchR) which regulates epidermal homeostasis. We first report that comparing human peripheral blood mononuclear cells (PBMCs) from 4 affected and 15 unaffected members from the family with MDM revealed that secretion of normal SLURP-1 protein and T-cell activation were impaired in PBMCs with the heterozygous and homozygous SLURP-1 G86R mutation. Addition of recombinant SLURP-1 protein would reconstitute T-cell activation in our MDM patients. 2 of 4 affected members developed melanoma. We also detected cancer stem cell markers by analyzing mRNA of PBMC in 3 of 3 MDM patients. Since there is currently no effective treatment for metastatic melanoma, identifying the novel molecular pathomechanisms is crucial for the development of new treatments.In this project, we will investigate the roles of SLURP-1 in aggressiveness of melanoma and MSCs in 3 years:The 1st year:(1) Comparing the stem cell markers, SLURP-1expression and nAchRs expression among tissue specimens of various staged melanoma.Comparing the level of SLURP-1 expression among various melanoma cell lines, ABCB5+ melanoma cells, and ABCB5- melanoma cells; Assess the biological function of the SLURP-1 protein in melanoma cell lines, ABCB5+ melanoma cells, and ABCB5- melanoma cells including proliferation, apoptosis, differentiation, tumorigenesis, melanosphere formation and nAchR signalling pathway.The 2nd year:(1) Investigate and verify the interaction between T-cells and melanoma cells / MSCs(2) Confirm the function of SLURP-1 in tumorigenesis, angiogenesis, metastasis and stem cell markers on xenotransplant models. (IL2Rγ-/- NOD SCID mice)The 3rd year:(1) Investigate the cancer biological effects of SLURP-1 silencing by using dominant-negative SLURP-1 in melanoma cell lines (A2058 and other cell lines), ABCB5+ melanoma cells, and ABCB5- melanoma cells (tumorigenesis, anti-apoptosis, angiogenesis , melanoma stem cells formation);(2) Investigate the effects of SLURP-1 overexpression in melanoma cells by transfection with SLURP-1 expression vector .The aim of this project is to investigate the roles and molecular mechanisms of SLURP-1 in melanoma carcinogenesis, which may be used to the development of novel treatments for melanoma.惡性黑色素瘤黑色素瘤幹細胞SLURP-1 基因Mal de MeledaMelanomaMelanoma stem cellsSLURP-1Mal de Meleda