鼻咽癌細胞上與樹突狀細胞的DC-SIGN 交互作用蛋白之鑑定

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題名:	鼻咽癌細胞上與樹突狀細胞的DC-SIGN 交互作用蛋白之鑑定 Identification of DC-SIGN-interacting proteins in nasopharyngeal carcinoma cells
作者:	賴政豪 Lai, Chang-Hao
貢獻者:	醫學檢驗暨生物技術學系陳建和林詠峯
關鍵詞:	鼻咽;癌細胞;樹突狀;細胞 DC-SIGN;nasopharyngeal carcinoma cells
日期:	2012-06-27
上傳時間:	2018-11-16 10:00:35 (UTC+8)
摘要:	鼻咽癌細胞是一種盛行於東南亞的惡性上皮細胞癌。目前已知在癌症的形成過程中，對免疫系統的躲避作用是一個重要的關鍵，然而鼻咽癌細胞是透過何種方式來躲避免疫系統監控目前尚未清楚。在人類免疫系統裡擔任重要抗原呈現角色的樹突狀細胞上有一種C型凝集素(C-type lectin)，樹突狀細胞特異性細胞間黏附分子-3-辨認非整合素(DC-SIGN)，它會辨認一些特殊的醣類結構並與其結合，然後誘發抑制型的免疫反應。研究指出癌細胞會表現許多不正常醣化蛋白，而這些醣化蛋白與免疫抑制有關。於是我們先用流式細胞儀以及螢光染色法偵測到鼻咽癌TW01細胞上確實有DC-SIGN可辨認的配體（ligand）;接下來再以細胞共同培養的方式，並用酵素免疫吸附法(ELISA）去檢測，也發現鼻咽癌細胞會刺激樹突狀細胞分泌免疫抑制的細胞激素，如介白素10（IL-10）;我們更進一步利用DC-SIGN抗體和微小干擾核苷酸(siRNA)去拮抗DC-SIGN的作用，發現IL-10的量明顯減少;接著用DC-SIGN重組蛋白以免疫沉澱法將鼻咽癌細胞上與DC-SIGN交互作用的膜蛋白萃取出來，並利用LC/MS/MS去鑑定。其中被鑑定出來的Annexin A2在TW01細胞株表現是主要的的蛋白之一。以上結果顯示，鼻咽癌細胞上表現DC-SIGN可辨認的ligand，並透過樹突狀細胞上的DC-SIGN刺激其分泌免疫抑制性的IL-10，進而躲避免疫系統的監控。另外，本研究中鑑定出來的Annexin A2，將可作為一個鼻咽癌細胞上的新標誌，未來可繼續研究Annexin A2 與樹突狀細胞之間的關係。 Nasopharyngeal carcinoma (NPC) is a malignant epithelial cell-derived tumor that is predominately found among populations in Southeast Asia, including Taiwan. The interaction that exists between tumors and the immune system is important in tumor progression. Because the mechanisms underlying NPC’s ability to escape the surveillance of the immune system are still unclear, our study sought to investigate the interaction between one type of antigen presenting cells, known as Dendritic cells (DCs), and NPC. DCs express a C-type lectin, DC-pecific ICAM-3-Grabbing Nonintegrin (DC-SIGN) that can induce suppressive immune responses by recognizing carbohydrate structure. Recent studies indicated that cancer cells express many abnormal glycosylated proteins, and these glycoproteins are involved in the immune suppressive responses. Therefore we used flow cytometry and immunostaining, and found that DC-SIGN-recognized ligands were expressed on NPC cells. Using enzyme-linked immunosorbent assay (ELISA), we demonstrated that NPC cells induced immunosuppressive interleukin (IL)-10 secretion from DC. We further showed that IL-10 secretion was inhibited by DC-SIGN antibody and siRNA. We immunoprecipitated the NPC membrane proteins using purified recombinant DC-SIGN protein, and identified the unknown proteins by LC/MS/MS. Annexin A2 was identified as a major protein which is highly expressed in TW01 NPC cells. In conclusion, NPC cells express DC-SIGN ligands, and stimulate immunosuppressive IL-10 secretion from DC through DC-SIGN activation, resulting in the escape from surveillance of the immune system. We are focusing on annexin A2, a novel target on nasopharyngeal carcinoma, and will research the relationship of annexin a2 and DC-SIGN in the future.
描述:	碩士指導教授-陳建和共同指導教授-林詠峯委員-黃嘯谷委員-廖辰中委員-鄭朝文
資料類型:	thesis
顯示於類別:	[醫學檢驗暨生物技術學系所] 博碩士論文

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