2012-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/657170摘要：膀胱尿路上皮癌 (urothelial carcinomas; UCs)是泌尿系統常見的惡性腫瘤，發生率逐年上升。轉移性的尿路上皮癌，需接受化學治療，但終會會因抗藥性而至病患死亡，這一直是臨床治療的重要課題。在癌細胞中有少量的細胞稱為腫瘤幹細胞 (Cancer stem cell；CSC)，和腫瘤生成、維持 (maintenance)及腫瘤進展 (progression)密切相關。此類細胞於體外經培養可形成未分化癌症次球體 (Cancer spheres)並且已經被報告與癌症轉移及化學抗藥性 (chemoresistance)有關。此為三年計畫第一年: 重點在建立多重抗藥性及腫瘤幹細胞的in vitro and in vivo研究平台。吾人已建立一株多重抗藥性的尿路上皮癌細胞 (NTUB1/R)，並初步確認其stemness 特性。將進一步建立尿路上皮癌腫瘤幹細胞 (CSCs)，並確認腫瘤幹細胞及多重抗藥性尿路上皮癌細胞的stemness markers 如Nanog, OCT4, SOX2.之表現，再生能力(Self-renew)、轉移能力及化學抗藥性。第二年：我們已初步證實在化學抗藥性的尿路上皮癌組織中，其stemness marker表現量增加。將進一步在臨床手術檢體確認stemness markers與腫瘤行為，化療反應及預後的相關性。此外，吾人將建立腫瘤幹細胞及多重抗藥性尿路上皮癌細胞的異體移植的動物研究模式，並將第一年的結果在in vivo中證實。第三年：評估stemness markers在癌症侵犯，化學抗藥性扮演的角色，利用downregulation, knockdown或overexpression的模式來證實其功能。我們的初步結果可見這些藥物確實對NTUB1/R有效。將利用一些新的治療藥物(Histone deacetylase (HDAC) inhibitor, DNA methylation Inhibitor, NEDD8 activating enzyme inhibitor)或合併現有的化療藥物(cisplatin; paclitaxel)能否加強化學治療的效果，改變UC細胞的抗藥性？這些藥物能否藉由調控stemness merker，造成UC細胞凋亡？能否reverse chemoresistance？在in vivo的動物模式中驗證。尿路上皮癌是一多重性的腫瘤，不同的腫瘤型態，有截然不同的預後及腫瘤行為，腫瘤幹細胞應在其中扮演重要角色。吾人相信，藉由此腫瘤幹細胞的研究，將能找到克服化學抗藥性的新的治療模式。<br> Abstract: Bladder urothelial carcinoma (UC) is a common malignancy and the incidence is increasing by years in Taiwan. Chemoresistance was inevitable in treatment of metastatic disease and lead to the ominous outcomes. To develop novel therapeutic strategies to overcome chemoresistance is imperative. Emerging data showed a rare subpopulation of cancer cells, termed cancer stem cells (CSCs), is capable of initiating, maintaining and expanding the growth of tumor. These cells grow in vitro as undifferentiated cancer spheres and may be responsible for metastasis and chemoresistance, which may be the chief target of chemoresistance. In the preliminary data, we have established a multidrug resistance UC cell line (NTUB1/R) which showed the expression of stemness markers. These stemness markers (LIN28A and SOX2) were also highly expressed in chemoresistant clinical specimens.This is a three-year project:In the first year, we aim to establish the platform of mutidrug-resistant cells and CSCs research. We have establish multidrug resistant UC cell line (NTUB1/R) and proved the stemness characteristics. We will cultivate the CSCs of UCs by spheroid cells formation assay and side population determination. In addition, we will prove the expression of stemness and EMT markers and examine the chemosensitivity and invasion ability in CSCs.In the second year, we aim to prove the expressions of stemness markers (OCT-4, Nanog, SOX-2 and LIN28A) in the surgical specimen (UCs and chemoresistant UCs) and correlated these expressions with clinicopathological and outcomes. Besides, we will establish xenograft animal model from CSCs and NTUB1/R to prove their tumorigenesis, invasiveness and chemoresistance; furthermore to clarify the roles stemness markers play in vivo.In the third year, we will further evaluate the mechnisms of stemness markers in tumor invasion, chemoresistance by downregulation, knockdown and overexpression model. Besides, we try to find out novel drugs to conquer chemoresistance by focusing on histone deacetylase (HDAC) inhibitor, DNA methylation Inhibitor, NEDD8 activating enzyme inhibitor).UCs are heterogenous tumors and have diverse behavior and outcomes. We believe that CSCs plays an important role in cancer progression, metastases, and chemoresistance. CSCs will offer novel ideas for the lab and the clinic research to design new strategies of conquering the chemoresistance of UCs.膀胱尿路上皮癌 (Bladder urothelial carcinoma)腫瘤幹細胞 (cancer stem cellCSCs)上皮細胞轉變成間葉細胞的過程 (epithelial-to-mesenchymal transitionEMT)化學抗藥性 （chemoresistance）Bladder urothelial carcinoma (UC)cancer stem cell (CSCs)epithelial-to-mesenchymal transition (EMT) and chemoresistance