台灣西南沿海烏腳病流行區因長期飲用含砷井水而導致此一地區是全世界膀胱癌發生率最高的地區。本研究希望利用液相層析串聯式質譜儀(HPLC-MS/MS)來分析膀胱癌尿液中之蛋白質，同時希望也藉此找出台灣與砷中毒有關膀胱癌之致癌機轉。我們收集了24位病人，其中有8例膀胱癌來自烏腳病流行區，8例膀胱癌來自烏腳病非流行區及8例正常人的尿液，經導尿方式取得尿液樣本。為了避免在質譜分析時大量的蛋白質會抑制少量蛋白質的訊號，因此將樣本利用acetonitrile沈澱法和分子篩去除尿液中的基質，再使用高效能液相層析電噴灑離子化質譜儀 (High performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry,HPLC-ESI-MS/MS) 分析尿液中蛋白質體。總共鑑定到146個蛋白質體，再使用生物資訊工具去篩選蛋白質名單，我們自與砷中毒有關膀胱癌患者尿液中分析出三種有意義的尿液蛋白質─ADAM 28, calpain 9 及 ring finger protein 20，有可能成為與砷中毒有關膀胱癌的腫瘤標誌；但是是否可成為真正新的膀胱癌腫瘤標記尚待進一步研究。HPLC-ESI-MS/MS可以一次大量分析尿液中蛋白質的質量及蛋白質的成份來鑑定烏腳病流行區膀胱癌尿液中蛋白質是否與其他非烏腳病流行區膀胱癌尿液中蛋白質組成是否有差異，進而探討烏腳病流行區與砷有關膀胱癌致病機轉。

Arsenic is widely distributed in nature and is mainly transported in the environment by water. Consumption of artesian well water with high levels of arsenic has been associated with genitourinary cancer, especially bladder transitional cell carcinoma (TCC). To search for biomarkers that are specific for arsenic associated with the diagnosis of bladder TCC, proteins in the urine of normal individuals and patients with either bladder TCC or arsenic-associated bladder TCC were systematically examined by High performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry,HPLC-ESI-MS/MS). Urine specimens were collected by catheterization from patients and age- and sex-matched healthy individuals. A HPLC-ESI-MS/MS was used to generate proteome patterns from urine specimens obtained from patients with As-associated (n=8) and non-As-associated (n=8) primary transitional cell carcinoma (TCC) and from sex- and age-matched healthy individuals (n=8). Three urinary proteins were found to have significantly altered levels in patients following chronic arsenic exposure. These proteins were a disintegrin and metalloprotease (ADAM) protein, calpain 9, and ring finger protein 20. The large-scale identification of urinary proteomes using HPLC- ESI-MS/MS may serve as an ideal and efficient method for the establishment of a panel of potential arsenic-associated TCC biomarkers and may help elucidate the mechanisms involved in bladder cancer induced by chronic arsenic exposure.

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