2010-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656849摘要：胃癌是全世界人口的的第一大健康威脅。在台灣地區也分居男性十大腫瘤的第四名與女性十大腫瘤的第六位。許多流行病學與動物模式的研究中早已確立胃癌的發生與幽門螺旋桿菌的感染是息息相關的，而國際癌症研究署(IARC)也於西元1994 年將幽門螺旋桿菌定義為第一類的致癌因子。目前發現世界上超過半數的人口會受到幽門螺旋桿菌的感染，受感染的個體中約有百分之十會發展成為嚴重的胃部疾病，例如：胃潰瘍與胃部發炎萎縮，只有百分之一會造成胃癌的發生。幽門螺旋桿菌的感染會造成如此歧異的致病結果，除了本身病毒因子的差異與環境因素的影響外，宿主本身的細胞因子也扮演了決定性的角色。過去本實驗室利用蛋白質體學技術，發現許多經幽門螺旋桿菌感染後表現量會上升的蛋白質，其中又以缬絡胺酸蛋白質(valosin-containing protein)這個重要蛋白質表現差異最大。Valosin-containing protein (VCP)的分子量為97kDa，是屬於第二型AAA (ATPase associated witha variety of activities) ATPase 的一員，它由六個相同次體組成的六具體 (homo-hexamer)，形成相疊的環狀構造，如同其它AAA 蛋白質，VCP 在細胞中可以催化ATP 水解以製造能量，並利用這些能量來執行工作。VCP 對細胞而言是個必要的蛋白質，它廣泛存在於細胞中，約佔細胞裡所有蛋白質的1%。VCP 在細胞中扮演各式各樣的角色，包括膜融合、蛋白質的降解和內質網相關的降解等功能的調控。此外已有許多研究指出，在不同種類癌症中，包括罹患非小細胞肺癌、肝癌、胰臟腺癌以及胃癌的病人腫瘤組織，其VCP 蛋白質表現量皆比正常組織高，因此，VCP 表現量的多寡，可以用來偵測癌症發展的生物指標。近年來發現 VCP 在許多不同的癌症組織都有大量表現的情形，且其在病人組織中的含量可作為癌症癒後狀況的指標。然而VCP 與癌症發展之間的關係目前尚未完全研究清楚，為了繼續探討VCP 在胃幽門螺旋桿菌致癌過程中所扮演的角色，我們希望結合蛋白質體學技術和免疫沉澱(immunoprecipitation)方法，分析與VCP 交互作用之蛋白複合體，進而瞭解該目標蛋白之生物功能。鑑定受胃幽門螺旋桿菌調控之細胞因子並分析其功能特性，不僅增加我們對於病原菌與宿主間的相互影響，更有助於發展新的胃癌治療方向。<br> Abstract: Gastric cancer remains a major health problem being the second commonest malignancy inthe world. In Taiwan, gastric cancer also stays the fourth-common cause of cancer death in male,and the sixth in female. Epidemiological and animal studies demonstrated a link between gastriccancer and chronic infection with Helicobacter pylori (H. pylori), and H. pylori infection has beenclassified in 1994 by the International Agency for Research against Cancer (IARC) as a carcinogentype I for gastric cancer. More than 50% world-wide population was infected by H. pylori, though,approximately 10% of the infected subjects will develop more severe gastric pathologies like pepticulcer disease and atrophic gastritis, and only 1% of infected individuals will development gastriccancer. In addition to H. pylori virulence factors and environmental influences, host factorsdetermine such divergent disease outcome.Previous reports have demonstrated that valosin-containing protein (VCP) is upregulated afterHelicobacter pylori infection, and is associated with H. pylori-induced gastric carcinogenesis.Valosin-containing protein (p97 or VCP) is a member of the type II AAA (ATP associated with avariety of activities) ATPase. VCP is ubiquitous, essential, accounting for more 1% of the totalcellular protein. It involves in an unusually wide variety of functions, such as membrane fusion,ubiquitin-proteasome mediated proteolysis, ER-associated degradation. In addition, several lines ofevident indicate that expression level of VCP is strongly associated with tumor progression andprognosis, including non-small cell lung cancer, hepatocellular carcinoma, pancereatic ductaladenocarcinoma, and gastric carcinoma. So the level of VCP in cancerous tissues is a prognosismarker for cancer diseases.However it is not a clear relationship between VCP expression level and cancer development.Therefore, we sought to evaluate the functional roles of VCP which may be involved in thepathogenesis of H. pylori-induced gastric diseases, proliferation and anti-apoptosis. We usedoverexpressing VCP in AGS cells and co-immunoprecipitating the whole complexes, further thecomplexes are identified by using proteomics approaches. Finally, we will be established and usedto investigate their signaling complexes and networks. The identification of such unknown host cellfactors will add to our understanding of host-pathogen interactions and might help in thedevelopment of new therapeutic strategies.胃癌缬絡胺酸蛋白質(VCP)幽門螺旋桿菌免疫沉澱gastric cancervalosin containing-proteinco-immunoprecipiationinteractomesignal transduction networks