臺灣博碩士論文加值系統

有越來越多的研究證據顯示，高度糖化終產物(AGE)及其受器(RAGE)與腫瘤的形成和轉移有著密切的關係。然而有關它們與特定部位癌症發生的關聯性研究，包含大腸癌，目前為止仍是相當少的。本論文研究將探討高度糖化終產物和RAGE是否與調控人類大腸癌細胞株(HT-29和SW-480)生長和細胞週期進行有關。我們的實驗結果發現，使用高度糖化終產物處理HT-29以及SW-480細胞後會呈現濃度和時間依賴性的效應，明顯促進細胞的生長，但是非糖基化白蛋白(BSA)處理後並沒有上述的效應產生。而大腸癌細胞在非糖基化白蛋白和高度糖化終產物處理下並沒有造成細胞毒性的改變。再者，我們發現在大腸癌細胞中，高度糖化終產物會使JAK2-STAT1訊息傳遞途徑顯著地活化。而此JAK2-STAT1活化程度以及細胞生長都會受到JAK2抑制劑AG490所調降。我們也進一步得知anti-RAGE抗體會減少高度糖化終產物促進的JNK和p38 MAPK蛋白質磷酸化，並阻斷高度糖化終產物所誘發之細胞生長。所以本篇研究結果顯示高度糖化終產物及其受器RAGE對大腸癌細胞生長有促進的作用，而JAK2-STAT1還有JNK-p38 MAPK訊息傳遞途徑或許可被當成是未來治療大腸癌的目標之一。

Mounting evidence indicates that advanced glycation end-products (AGE) and their receptors (RAGE) have been implicated in tumor development and metastatic progression. However, prospective data on their association with cancer of specific sites, including colorectal cancer, are limited. We examined here whether and how AGE/RAGE could regulate cell growth in human colorectal adenocarcinoma cell lines (HT-29 and SW-480). We found that exposure of HT-29 and SW-480 cells to AGE (but not non-glycated BSA) increased the cell growth in concentration- and time-dependent manners. There were no changes in cytotoxicity in BSA or AGE treatment in these cells. Moreover, the JAK2-STAT1 signaling pathway was markedly activated by AGE in human colorectal cancer cells. The specific JAK2 inhibitor AG490 significantly prevented the proteins phosphorylation of JAK2 and STAT1 and reduced the effect of AGE-accelerated cellular growth. We also found that anti-RAGE antibody decreased the phosphorylation of JNK and p38 MAPK proteins and blocked the effect of AGE-induced cellular proliferation. These results suggest that AGE/RAGE has potent stimulatory effect on colorectal cancer cell growth, and the JAK2-STAT1 and the JNK-p38 MAPK signaling pathways can thus be considered as possible targets for colorectal cancer therapy.

口試委員審定書--------------------------------------------------------------------------------i
授權書--------------------------------------------------------------------------------------ii
致謝---------------------------------------------------------------------------------------iii
目錄----------------------------------------------------------------------------------------iv
圖表目錄-------------------------------------------------------------------------------------vi
英文摘要-----------------------------------------------------------------------------------viii
中文摘要-------------------------------------------------------------------------------------ix
縮寫檢索表------------------------------------------------------------------------------------x

第一章 緒論--------------------------------------------------------------------------------1-11
1. 大腸癌治病機轉-----------------------------------------------------------------------------2
2. 糖與癌細胞之間的關係------------------------------------------------------------------------3
3. 高度糖化終產物的形成------------------------------------------------------------------------3
4. JAK-STAT訊息傳遞路徑-----------------------------------------------------------------------5
5. MAPK訊息傳遞路徑---------------------------------------------------------------------------7
6. 細胞週期的調控-----------------------------------------------------------------------------9
7. 研究目的----------------------------------------------------------------------------------11

第二章 材料與方法---------------------------------------------------------------------------12-20
1. 細胞培養技術-------------------------------------------------------------------------------13
2. 細胞計數----------------------------------------------------------------------------------14
3. 高度糖化終產物(AGE)之製備-------------------------------------------------------------------15
4. 細胞增生分析法-----------------------------------------------------------------------------15
5. 細胞毒性試驗；乳酸去氫酶分析法----------------------------------------------------------------16
6. 硫酸十二酯納-聚丙烯醯胺凝膠電泳---------------------------------------------------------------17
7. 西方墨點分析-------------------------------------------------------------------------------18
8. 細胞溶解物之製備----------------------------------------------------------------------------19
9. 細胞蛋白質總量之測定-------------------------------------------------------------------------19
10. 統計分析----------------------------------------------------------------------------------20

第三章 結果--------------------------------------------------------------------------------21-25
第四章 討論--------------------------------------------------------------------------------26-30
第五章 圖表--------------------------------------------------------------------------------31-41
第六章 參考文獻----------------------------------------------------------------------------42-47

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