胃癌早期檢測之新穎性血漿刀豆素A吸附醣蛋白生物標記的開發計畫

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Title:	胃癌早期檢測之新穎性血漿刀豆素A吸附醣蛋白生物標記的開發計畫 The development plan of plasma novel Con A-bound glycoprotein biomarkers for early detection of gastric cancer
Authors:	李冠緯 Lee, Kuan-Wei
Keywords:	胃癌;刀豆素A;gastric cancer;concanavalin A
Date:	2012-07-06
Issue Date:	2018-11-16 11:02:40 (UTC+8)
Abstract:	在台灣胃癌的死亡率是居癌症死因的第六位，然而當胃癌在癌症早期就被診斷出來時，其生存率高達95%以上，故一個能早期診斷胃癌的生物標記是迫切需要的。在本研究中，我們使用刀豆素A (concanavalin A, Con A)親和層析法、一維凝膠電泳和奈米級液相層析串聯質譜從血漿樣品中篩選候選生物標記，血漿樣品來自30位胃癌患者和30位健康受試者。第一步，我們以年齡及性別配對混合血漿樣品，從中鑑定出22個Con A吸附表現量有差異的蛋白質，其中包括12個表現量上升的蛋白質和10個表現量下降的蛋白質，經學生t檢定，p<0.05。並且，有兩個表現量上升的蛋白質其表現量差異達三倍以上， 1個是leucine-rich alpha-2-glycoprotein (LRG1)，1個是inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3)，此兩個蛋白質將以西方墨點法分析驗證其表現量的差異。接著，我們以Ingenuity Pathway Analysis (IPA)軟體分析這22個表現量有差異的蛋白質功能，分析的類別有功能分析(Functional Analysis)、典型的路徑分析(Canonical Pathway Analysis)和網路(network)，其中網路包括發炎反應和細胞與細胞之間的信號及相互作用；典型的路徑分析包括急性期反應信號、補體系統、凝血系統、原發性免疫缺失信號和系統性紅斑性狼瘡信號。我們的結果認為Con A吸附LRG1和ITIH3可能是具潛力可以作為胃癌早期檢測的血漿生物標記。除此之外，我們在ITIH3上找到三個新穎性的轉譯後修飾：asparagine-41的hexose-N-acetyl-hexosamine (HexHexNAc)、aspartic acid-290的trimethylation和histidine-335的flavin adenine dinucleotide (FAD)。 Gastric cancer is the sixth leading cause of cancer deaths in Taiwan, yet survival rates are over 95% when it is diagnosed at an early stage, emphasizing the need for biomarkers for early diagnosis. In the present study, we used concanavalin A (Con A) affinity chromatography, one-dimensional gel electrophoresis and nano-LC-MS/MS to screen biomarker candidates in plasma samples obtained from 30 patients with gastric cancer and 30 healthy volunteers. First, we pooled the plasma samples matched by age and sex status. We identified 22 differentially expressed Con A-bound glycoproteins including 12 up-regulated proteins and 10 down-regulated proteins; these differences were significant (Student’s t-test, p-value less than 0.05). Furthermore, two up-regulated proteins had an expression level that was three-fold or greater in gastric cancer samples when compared with normal control samples; these proteins included leucine-rich alpha-2-glycoprotein (LRG1) and inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3), and the expression levels were validated by Western blot analysis. Functional analysis of the differentially expressed proteins with Ingenuity Pathway Analysis reveals vital networks including inflammatory response, and cell-to-cell signaling and interaction and canonical pathways involving acute phase response signaling, complement system, coagulation system and primary immunodeficiency signaling. Our results suggest that Con A-bound LRG1 and ITIH3 may be potential plasma biomarkers for the early detection of gastric cancer. In addition, three novel post-translational modifications in ITIH3 were identified: hexose-N-acetyl-hexosamine (HexHexNAc) at asparagine-41, trimethylation at aspartic acid-290, and flavin adenine dinucleotide (FAD) covalently bound at histidine-335.
Description:	碩士委員-陳建仁委員-陳仲瑄共同指導教授-廖辰中指導教授-林景堉
Data Type:	thesis
Appears in Collections:	[ ] Dissertations/Theses

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