2011-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/655338摘要：Glucose regulated protein 78（GRP78）在細胞內扮演著molecular chaperone的角色，為維持細胞內恆定性的蛋白，能調控內質網壓力（ER stress)引發之細胞凋亡。少數研究已經證實GRP78在許多惡性腫瘤有高度表現，並且與化療抗藥性及預後有關。GRP78在泌尿上皮癌（UC）的表現，在過去並無相關研究。而我們初步結果顯示出UC腫瘤之GRP78表現量較高，GRP78高表現的病人預後較差，調控ER stress能造成UC 細胞凋亡，我們發現兩種藥物，2-methoxyestradiol (2-ME) 及 celecoxib可藉由調控ER stress路徑，造成UC細胞凋亡。此為三年計畫第一年：吾人將利用臨床上收集之泌尿上皮癌之組織，研究其GRP78表現，及與病人之臨床病理資料及預後之相關情形？能否作為預後因子？利用不同泌尿上皮癌的細胞株及已建立的的抗藥株，偵測GRP78表現差異與腫瘤抗藥性及侵犯性間的關係。第二年：建立GRP78 overexpressing及knockdown之泌尿上皮癌細胞株，來進一步證實GRP78與腫瘤侵犯及chemoresistance的相關性？調控ER stress是否會改變chemoreistance，造成UC細胞凋亡？利用調控ER stress藥物合併化療藥物能否加強治療的效果？口服藥物2-ME及celecoxib能調控ER stress下游機轉 (PERK, IRE1或ATF6 pathway)，造成UC細胞凋亡。藥物合併ER stress之調控之治療效果?第三年：根據建立之實驗模式，可用以測試其他的molecular chaperones (GRPs, Hsps, ORPs等), 找尋新的腫瘤治療模式。吾人也將建立in vivo tumor heterotransplanted之動物模式，在BALB/c小鼠皮下上植入轉殖UC細胞株。在動物模式下，進一步證實2-ME及celecoxib在調控ER stress的機轉。化療藥物在不同GRP78表現下之活體治療效果。<br> Abstract: We encountered several problems in treating urothelial carcinoma (UC). First, there still no biomarkers for clinical use to predict tumor behavior. Second, chemoresistance is inevitable in metastatic disease. Exploring novel anti-cancer modalities to conquer drug resistance in UC is imperative. Glucose regulated protein 78 (GRP78), a main molecular chaperone, plays an important role in cancer biology and endoplasmic reticulum (ER) stress. The associations between GRP78 expression and cancer behavior, chemoresistance have been confirmed in various cancers. However, relevant studies in UC have never been reported. In our preliminary data, we demonstrated high level of expression in NTUB1 cells and UC tissues. The expression was significantly associated with prognosis. We also identified two oral drugs, 2-methoxyestradiol (2-ME) and celecoxib, which could induce apoptosis by regulation ER stress pathways.This is a three-year project.In the first year, we will examine the expression of GRP78 in UC cells and tissues. We aim to clarify the correlations between GRP 78 expression and clinicopathological parameters. Besides, we want to investigate the expression of GRP78 in UC cell lines and drug-resistant sublines to clarify the associations between GRP78 expression and tumor invasion ability as well as chemosensitivity in vitro.In the second year, we aim to establish the GRP78 overexpressing and knockdown model in vitro. The associations between GRP78 expression and cell proliferation, invasion ability and chemosensitivity will be explored. Whether the regulation of ER stress has effect on chemoresistance. Exploring down-stream mechanisms of 2-ME (PERK, IRE1, or ATF6 pathway) . By regulation of ER stress (knocking down GRP78 combined with 2-ME or overexpressing GRP78 combined with celecoxib) , can we potentiate cytotoxic effect or reverse the chemoresistance of in UC cells?In the third year, we will establish the in vivo heterotransplanted model by giving an subcutaneous injection of NTUB1 cells. We try to further prove the cytotoxic mechanism of 2-ME or celecoxib via ER stress regulation in vivo. We also aim to test whether modulation of GRP78 would affect the in vivo chemosensitivity of UC cells in animal model. On the basis of the model we established, we could further examine other stress-associated molecular chaperones such as GRP94, 170, Hsp 28, 60, 70, 90, ORP 150 to develop new modalities of UCs therapy.泌尿上皮癌內質網壓力GRP782-methoxyestradiolcelecoxib預後細胞毒殺作用化學治療反應Urothelial carcinoma2-methoxyestradiolcelecoxibendoplasmic reticulum stressglucose regulated protein 78 (GRP78)prognosischemoresistance