膀胱中的上皮組織是屬於一種移形上皮細胞，它就像其它腸胃道的表皮細胞，會不斷持續地分裂生長、老化、掉落，台灣地區所罹患的泌尿系統癌症大部分以膀胱移形上皮細胞癌（Transitional Cell Carcinomas, TCC）為主，膀胱移形上皮細胞癌的發生就是因為移形上皮組織不正常的分裂增殖所導致的疾病，膀胱癌中有高達90％為移形上皮細胞癌，其發生率有逐年增加的趨勢。由於TCC發展緩慢，大部分的病人於診斷發現TCC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，但只要能早期發現，病人的存活率很高。
有鑑於此，傳統的TCC鑑定方法從診斷到判定以何種治療方式及決定癌症的分期和病人的存活率時，可能已錯過治療的黃金時機；因此，為了能降低TCC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。
為了能開發出早期診斷TCC的系統，本論文以蛋白質組學技術得到台灣病人TCC細胞株整體基因表現情形，探討在不同級的TCC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TCC的早期腫瘤標記（tumor marker）。我們使用蛋白質二維電泳並利用靈敏度高的銀染法呈色，取得不同級TCC細胞株的二維電泳圖譜，將所得到的圖譜進一步以軟體做分析比對，找出具有表現差異的蛋白質點，並利用串聯式質譜儀（LC/MS/MS）進行序列比對，鑑定出這些蛋白質的身份。
我們使用兩株TCC細胞株：TSGH-8301 (grade II)、BFTC905 (grade III)細胞株。利用蛋白質體學技術找出5個有表現差異的蛋白質點，分別是prohibitin、maspin、S100A4及hsp27的蛋白質。其中prohibitin及maspin為tumor suppressor gene，而S100A4的蛋白質於目前的文獻報導中指出在乳癌及膀胱癌細胞中若有此蛋白質存在下，癌細胞具有較高度的轉移性（metastasis）。兩個相同分子量但不同pI的點均被鑑定為hsp27，此現象可能與轉譯後修飾有關。在TSGH-8301細胞株中prohibitin、maspin及hsp27產生量比BFTC905細胞株高，這個結果顯示TSGH-8301細胞株因有較高量的tumor suppressor protein，可遏止因生長訊息傳遞失控而導致癌細胞不正常分化，使病患病理分期不會進展成Grade III的細胞；S100A4的蛋白質被發現只存在BFTC905細胞株中而不存在於TSGH-8301細胞株，此結果與BFTC905細胞株為較晚期的細胞是一致的。本論文找到五個表現差異的蛋白質對解開膀胱癌致癌機轉有所助益，但是否能成為膀胱癌早期診斷的分子符號則須在進一步的研究。 The epithelial tissue of bladder belongs to the transitional epithelial lining. Like the intestines and stomach path epithelial lining, the transitional epithelial cell of the bladder keeps going on living、aging and falling off the bladder. Formation of transitional Cell Carcinomas（TCC）attributes to the abnormal proliferation of the transitional epithelial cells. In Taiwan, approximately 90％ of bladder tumors are classified as TCC. The TCC occurrence rate increases every year and is the thirteenth leading cause of the mortality.
Due to the slow development of TCC, using the traditional TCC 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 TCC death rates during early diagnosis and prompt treatment.
In this study, the proteome maps of the TCC cancer cells of the different grades were established using proteomics method. In addition, the differentially expressed proteins during the TCC progression were found with the comparison between the proteome maps of the TCC cancer cells of different grades in hope that these proteins can be utilized as the tumor makers for the early detection. 2-D gel maps of two TCC cells of different grades (TSGH8301, grade II; BFTC905, grade III) were imaged and compared with PDQUEST analytical software and the differentially expressed protein spots were identified using electrospray ionization mass spectrometry (ESI/MS/MS).
Five differentially expressed protein spots were identified, which were prohibitin (tumor suppressor gene)、maspin (tumor suppressor gene)、S100A4 (metastasis-related gene) and heat shock protein 27 (hsp27). Two protein spots with the same molecular weight and different pI represented the hsp27 probably due to the translational modification. In TSGH-8301 cell line prohibitin、maspin and hsp27 were present at higher amount than in BFTC905 cell line, suggesting that the development of tumors possibly was prevented in the presence of higher amount tumor suppressor proteins and therefore reduce grade of TCC clinicopathological features were observed. Our results demonstrated that S100A4 protein was present only in the BFTC905 cells but not in TSGH8301 cells. Previous studies have demonstrated that S100A4 was found at higher amount in metastatic human breast and bladder tumor cells. This result was consistent with the fact that BFTC905 cell was the higher-grade cancer cell. In this study, five differentially expressed proteins were identified and may help to elucidate the TCC progression. In the future, it may be worthwhile to examine if these protein are suitable for being the tumor marker for early detection.
