Transmembrane and coiled-coil domains 1 (TMCO1)是一種穿膜蛋白，目前對於TMCO1的功能仍然未知。根據本實驗室先前的生物資訊學及史丹福微陣列資料庫 (Stanford Microarray Database, SMD)的探勘(data-mining)與初步的分析數據發現TMCO1，並利用IHC (immunohistochemistry) 發現TMCO1在非侵入性的膀胱癌具有高度的表現，因而想更進一步的研究TMCO1在膀胱癌中扮演的角色。首先，分析TMCO1 mRNA的內生性表現，發現在膀胱癌細胞株中表現量較正常細胞低，接著我們高度表現TMCO1基因，發現會抑制細胞增生、遷移、侵犯及非貼附性生長等能力，並使細胞週期停滯在G0/G1期；相反的，若抑制TMCO1基因表現會促進細胞遷移、侵犯及非貼附性生長之能力。此外，我們也發現過度表現TMCO1會減少AKT1活化與MDM2的蛋白質表現量，使核內TP53蛋白質表現量增加。在探討TMCO1磷酸化位點的實驗中發現，點突變TMCO1 S60A會增加AKT1的活化及減少TP53蛋白的表現量，使細胞週期停滯的現象消失。綜合以上結果，TMCO1在膀胱癌中扮演著腫瘤抑制基因的角色。

The product of transmembrane and coiled-coil domains (TMCO1) gene is a member of several eukaryotic protein with unknown function. In a preliminary Bioinformatics and Stanford Microarray Database data-mining and some of our preliminary data, immunohistochemistry (IHC) analysis identified TMCO1 was up-regulated in non-invasion bladder cancer. In this study, we explored the functions and regulatory mechanisms of TMCO1 in bladder cancer. Our results showed that TMCO1 mRNA expression was significantly reduced in bladder cancer cells compared to normal counterparts. Overexpression of TMCO1 in bladder cancer cell lines induced G0/G1 cell cycle arrest, inhibited cell viability, proliferation, migration, invasion and anchorage-independent cell growth. In contrast, downregulation of TMCO1 accelerated cell migration, invasion and anchorage-independent cell growth. Furthermore, we showed that overexpression of TMCO1 increased nuclear TP53 protein expression by reduced AKT1 activity and MDM2 expression. To further investigate TMCO1 phosphorylation, we found TMCO1 S60A mutant promote cell cycle recovery from G0/G1 arrest through the activation of AKT1 and decrease of TP53 expression. Taken together, our data indicated that TMCO1 functions as a tumor suppressor in bladder cancer by increasing TP53 expression.

論文審定書 i
致謝 ii
中文摘要 iii
英文摘要 iv
英文縮寫表 v
圖次 vii
表次 viii
壹、緒論 (Introduction) 1
貳、實驗材料與方法 (Materials and methods) 5
參、結果 (Results) 21
肆、討論 (Discussion) 37
伍、參考文獻 (References) 39

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