2009-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/654504摘要：膀胱癌是國人相當常見之腫瘤，其已知之生成原因，與化學物質(包括染劑以及砷)之污染有相當關聯，但其造成膀胱細胞癌化之機制仍不明瞭。由於生物資訊學以及癌細胞蛋白質體學之進展，許多與膀胱癌相關之基因標記逐漸浮現，其中鈣網酪蛋白calreticulin被發現與膀胱癌之生成極具相關性。為了解鈣網酪蛋白於膀胱癌細胞中之作用，本實驗室已成功構築表達低量鈣網酪蛋白之穩定膀胱癌細胞株- J82-CalRNAi。初步結果顯示，與對照組相比較，降低鈣網酪蛋白之表達會抑制膀胱癌細胞株之生長以及移行，並對細胞附著於基質之能力有重要之影響。此外，血管生成因子VEGF-A之表達亦受到降低鈣網酪蛋白之影響。降低鈣網酪蛋白之表達亦會抑制膀胱癌細胞株於裸鼠中形成腫瘤，並明顯降低肺臟及肝臟之轉移。因此我們認為鈣網酪蛋白很可能是影響膀胱癌細胞癌化之重要調控因子。本計畫之實驗目的，即為驗證鈣網酪蛋白為造成膀胱癌細胞轉移之重要因子之假設。我們計劃以核酸晶片以及蛋白質質體之手段，尋找受到鈣網酪蛋白調控之基因；並利用所建立之裸鼠模式，釐清鈣網酪蛋白於膀胱癌細胞附著以及轉移之機制。本計劃之研究成果，應可對膀胱癌病人未來之治療方式提供更多線索。<br> Abstract: Bladder cancer is one of the top leading causes of cancer related death world wide and in Taiwan. Calreticulin is a multifunctional calcium binding protein. Increase calreticulin expression in various cancers has recently been reported. In addition, urinary calreticulin has been suggested to be a potential marker for bladder cancer. However, the roles of calreticulin in tumor development are not clear. In order to clarify the roles of calreticulin in bladder cancer, we have successfully generated a bladder cancer cell line stably transfected with shRNA of calreticulin (J82-CalRNAi). Compared to control vector transfected cell, J82-CalRNAi has a lower proliferation rate, migrate slower and also adhere poorly to type-I collagen. These results suggested that alteration of the levels of calreticulin expression in cancer cells might affect bladder tumor progression. The expression levels of VEGF-A, an important regulator for angiogenesis, were also inhibited in the cell lines, suggested that the angiogenic behavior of the cancer cells might also regulated by this protein. In addition, J82-CalRNAi cell generated a smaller tumor in nude mice when compare to its vector control counter part. Most importantly, the J82-CalRNAi derived tumor in nude mice has much less sites of metastasis in lung and liver. These results strongly suggested that the expression patterns of calreticulin may be a marker for bladder cancer and our model also provide an excellent tool to further investigate the roles of calreticulin in bladder cancer development. In this proposal, we will first attempt to utilize our developed system to further clarify the down-stream genes regulated by calreticulin through DNA arrays and proteomic procedures. In the second part of the proposal, we will attempt to investigate the mechanism of CRT on bladder cancer cells adhesion and the relationships between bladder cancer cell adhesion and metastasis by using the nude mice model we have successfully developed.膀胱癌鈣網酪蛋白癌細胞轉移癌細胞附著血管生成Bladder cancercalreticulinmetastasisadhesionangiogenesis