- 學位論文
探討一系列人類泌尿上皮細胞癌之化學抗藥性機制
Characterization of Chemoresistant Mechanisms in A Series of Human Urothelial Carcinoma Cells
摘要
中文摘要 台灣地區所罹患的泌尿系統癌症大部分以泌尿上皮細胞癌（Urothelial Carcinomas, UC）為主。由於UC發展緩慢，大部分的病人於診斷發現UC時，其癌細胞可能已具有侵犯深層組織及轉移至遠處器官的能力。因此，為了能降低UC的死亡率，迫切急需能有一套完整的系統，能早期診斷做出更有效的治療與防治，以期能降低膀胱癌對國人的威脅。膀胱癌在臨床上多採用外科手術切除，之後再併用化學藥物或放射線治療；但是許多研究發現，其中大約有三分之一的病人會對化學藥物治療產生抗藥性 (Drug resistance)，造成治療與癒後上相當程度的瓶頸。於是，我們決定針對膀胱癌細胞株，對其抗藥性做分析。首先，我們取了之前實驗室所建立的具抗藥性之五株膀胱癌細胞，此五株細胞株是分別對臨床上常用的五種抗癌藥物：Doxorubicin (adriamycin)、Cisplatin、Gemcitabine、Paclitaxel (Taxol)與Arsenic trioxide具抗藥性。利用蛋白質體學(Proteomics)技術，探討在不同抗藥性之膀胱癌細胞株中，有那些蛋白質受到影響而有表現量的變化，藉以尋找膀胱癌之抗藥機轉。我們使用蛋白質二維電泳(2-dimensional electrophoresis)並利用靈敏度高的銀染法(Silver staining)呈色，取得不同抗藥性之膀胱癌細胞株的二維電泳圖譜，找出具有表現差異的蛋白質點，並利用串聯式質譜儀（LC/MSMS）進行分析，並運用MASCOT MS/MS Ions Search鑑定出這些蛋白質的身份。利用蛋白質體學技術找出24個有表現差異的蛋白質點，而我們先對Rac1蛋白質做一探討，Rac1膜蛋白於目前的文獻報導中指出，它是Rho家族的成員之一，Rac1除了會啟動細胞的生長、抗細胞凋亡的功能，而且也會藉由一些轉錄因子來調控基因的表現。故本論文找到二十四個表現差異的蛋白質，對解開膀胱癌抗藥性機轉有所助益，但是否能成為具抗藥性之膀胱癌診斷的分子標記則須再進一步的研究。
並列摘要
Abstract Formation of Urothelial Carcinomas (UC) attributes to the abnormal proliferation of the transitional epithelial cells. In Taiwan, approximately 90% of bladder tumors are classified as UC. The UC occurrence rate increases every year and is the thirteenth leading cause of the mortality. Due to the slow development of UC, using the traditional UC identification method to confirm the cancer stage, the therapy, and the survival rate, we might miss the best timing for treatment. Hence, we may have to find new system and reduce UC death rates during early diagnosis and prompt treatment. In this study, the proteome maps of the UC cancer cells of the different grades were established using proteomics method. In addition, the differentially expressed proteins during the UC progression were found with the comparison between the proteome maps of the UC cancer cells of different grades in hope that these proteins can be utilized as the tumor makers for the early detection. Bladder cancer is one of the most common malignancies occurring worldwide. Surgery and intravesical chemotherapy are the most common treatments for superficial bladder cancer. Although initial response rates of 70% can be achieved, most patients will have a recurrence. Resistance to chemotherapeutic drugs is the major problem that causes therapy failure. Therefore, the characters of drug resistance in bladder cancer cell lines were analyzed in this study. First, we used 5 cell-lines that have been drug resistant by Doxorubicin (Adriamycin), Cisplatin, Gemcitabine, Paclitaxel (Taxol) and Arsenic trioxide. In this study, we use “Proteomics” to find different proteins in the drug resistance cell-line. We use 2-dimentional electrophoresis and silver staining to get a lot of spots. We examined these spots by “LC/MSMS” and “MASCOT MS/MS Ions Search” to identify them. In the present work, 24 differentially expressed proteins were identified and may help to elucidate the TCC progression. First, we focus on “Rac1” membrane protein. Previously, some scientists think it is one of Rho family. Rac1 is capable of promoting cell survival, anti-apoptosis and regulating some transcription factors. In the future, it may be worthwhile to examine if these proteins are suitable for being the tumor marker for early detection.
參考文獻
延伸閱讀
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