抑制 RAS/ROCK 或 mTOR 對於人類尿路上皮癌的侵襲和轉移

最近的文獻指出，在全世界中泌尿道上皮細胞癌(Urothelial Carcinoma,UC)的發生率不斷地攀升，而UC中最常見的為膀胱癌，且以依據衛生署統計，得膀胱癌的發生率排名第14名，且年齡發率有趨於老人化，目前可以知道治療癌症以標靶治療的副作用相對其他治療方式較小，可以知道mTOR 為PI3K/AKT/mTOR中扮演一個重要的角色。本研究旨mTOR抑制劑 (everolimus) 對於膀胱癌的機制澄清，在小鼠是否能抑制特發性膀胱癌細胞的生長。實驗方法在細胞實驗部分，先培養T24、TSGH、HT1376、HT1197不同之膀胱癌細胞增生，再使用everolimus觀察對其抑制的作用與相關機制，使用transwell小型遷移實驗觀察everolimus或RAS抑制劑 (ROCK Inhibitor Y27632) 對於膀胱癌細胞移動性分析，再給予藥品用Electric Cell-Substrate Impedance Sensing (ECIS) 來觀察膀胱癌細胞之傷口癒合試驗。結果指出當給予藥品everolimus處理過後，T24、HT1376、HT1197細胞之mTOR、NFκB、MMP及p-GSK蛋白表現量有所下降，在TSGH細胞，蛋白表現量卻無明顯下降，而當給予Y27632處理過後的T24、HT1376、HT1197細胞的並無RAS/ ROCK的抑制，而在TSGH細胞的RAS/ ROCK蛋白卻有所抑制。小型遷移實驗，T24、HT1376、HT1197細胞使用everolimus和TSGH細胞使用Y27632遷移的表現量都有明顯下降。在動物實驗方面，將T24細胞使用everolimus 和TSGH細胞使用Y27632分別先做前處理過後，使用BALB/c Nude mice，將處理過之膀胱癌細胞種於皮下，每週觀察之體重及腫瘤大小。結果顯示當有給予標靶藥物之癌細胞，體重方面雖無太大變化，但腫瘤大小卻有顯著的減小。本實驗證實表明，在膀胱癌四種不同的細胞株是由不同的信號通路調控。經由通過確認癌細胞的分子信號通路方可改變標靶療法藥物才可正確的對症下藥。

Recent literature has reported that urothelial carcinoma (UC) is with continuously rising of incidence worldwide. Among UC, bladder cancer is the most common cancer. The incidence of age have tended to be olderer. Currently side effects of cancer treatment of a relatively small targeted therapy other treatments.mTOR is important role in PI3K/AKTmTOR pathway. That mTOR inhibitor (Everolimus) could inhibit the growth of idiopathic bladder cancer.However,the effect is limited of mTOR inhibitor for bladder cancer. This study examined everolimus for the heterogeneous mechanisms of bladder cancer need to be clarified. The T24, HT1376 and HT1197 cells after treatment with everolimus, mTOR, NFκB, MMP and p-GSK were decreased,whereas T24,HT1376 and HT1197 cells treated with Y27632 (Ras/ROCK inhibitor) Ras/ROCK is not be inhibited, Y27632 attenuated the Ras and related proteins of TSGH cells,but not in T24, HT1376, HT1197 cells. Using Transwell assay, the T24, HT1376, HT1197 cells amount of migration exposed to Everolimus and TSGH cells migrated numbers treated with Y27632 were dose-dependently decreased respectively. Using ECIS assay, the T24, HT1376, HT1197 cells amount of wound healing exposed to Everolimus and TSGH cells wound healing exposed treated with Y27632 were dose-dependently decreased respectively. Our study revealed that four different cell lines from human urothelial cell carcinoma (UCC) are regulated by different signaling pathways. In the future, the target therapy through confirming the molecular change of cancer cell firstly might increase to suit the medicine to the illness in patients.