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52 巻 (2008)
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52 巻, 2 号
選択された号の論文の5件中1～5を表示しています
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シンポジウム：臨床プロテオミクスの現状と将来像―網羅的から選択的解析へ／定性から定量的解析へ―
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斉藤 邦明, 竹田 真由, 山本 康子, 舩渡 忠男, 高橋 香奈子, 帖佐 光洋, 竹村 正男, 清島 満原稿種別: シンポジウム1：臨床プロテオミクスの現状と将来像―網羅的から選択的解析へ／定性から定量的解析へ―
2008 年 52 巻 2 号 p. 15-18
発行日: 2008年
公開日: 2009/12/04
ジャーナル フリーAlthough the restricted dynamic range of conventional proteomic technology using two-dimensional gels and mass spectrometry has limited applicability (i.e. plasma proteome), our recent studies of proteomics technologies using electrophoresis to discover and identify new biomarkers are summarized. Low-abundance proteins are rarely seen on traditional two-dimensional gel electrophoresis (2-DE) analysis of plasma because of the highly abundant soluble proteins such as albumin and globulin in plasma. In this study, serum proteins can be separated into two fractions by the Con A-Sepharose column (retained fraction was enriched in N-glycosylated proteins), which is used for the exclusion of albumin and is a simple method to analyze proteins in plasma. The % area of each protein band was compared between the different groups and proteins in each band were identified using LC/MS/MS. In addition, we also summarized recent results from focused brain proteomics to identify proteins associated with dysmnesia using 2-DE. Neuronal protein expression in viral infected mice was compared with that in non-infected mice using the Image Master 2D. We detected approximately 700 protein spots, of which approximately 40 spots were distinguishable between non-infected and virus-infected mice. This proteomic analysis is a useful tool for identifying proteins in mouse diseases model (especially using KO or TG mice).
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下重 美紀, 佐藤 礼子, 本田 一文, 尾野 雅哉, 山田 哲司原稿種別: シンポジウム1：臨床プロテオミクスの現状と将来像―網羅的から選択的解析へ／定性から定量的解析へ―
2008 年 52 巻 2 号 p. 19-23
発行日: 2008年
公開日: 2009/12/04
ジャーナル フリーβ-Catenin is a downstream effector of the Wnt signalling pathway and exerts its oncogenic activity by transactivating the target genes of TCF4 (T-cell factor-4). The aberrant transactivation of these TCF4-regulated genes by β-catenin plays a crucial role in early colorectal carcinogenesis. Recently, we identified that splicing factor-1 (SF1) was one of the proteins whose expression is regulated by the β-catenin / TCF4 complex using quantitative differential proteomics analysis (isotope-coded affinity tagging and mass spectrometry). SF1 negatively regulates β-catenin-evoked gene transactivation and cell proliferation. In addition, we found that SF1 was essential for the induction of alternative splicing by the β-catenin / TCF4 complex, and induced known cancer-related alternative by spliced variants, such as Wnt-induced secreted protein-1v and fibroblast growth factor receptor-3-ATII. In intestinal tissues, the expression of SF1 was found to be correlated with the differentiation status of intestinal epithelial cells and inversely correlated with tumorigenesis. To analyze the biological involvement of SF1 in the process of carcinogenesis in vivo, we developed SF1-deficient mice by gene trapping. Azoxymethane (AOM), a known carcinogen organotropic to the colon, was administered into SF1+/+ and SF1+/− mice. The average number and volume of colon tumors per mouse were significantly greater in SF1+/− than in SF1+/+ mice. The enhanced susceptibility of SF1 haploinsufficiency to AOM-induced colon tumorigenesis may be attributable to the suppressive role of SF1 in the Wnt signalling pathway.
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山田 尚之, 窪田 和幸, 河上 麻美, 中山 聡, 鈴木 榮一郎原稿種別: シンポジウム1：臨床プロテオミクスの現状と将来像―網羅的から選択的解析へ／定性から定量的解析へ―
2008 年 52 巻 2 号 p. 25-30
発行日: 2008年
公開日: 2009/12/04
ジャーナル フリーGenomics and proteomics have been useful in many different areas of medicine and health science as an aid to discover new biomarker for disease diagnosis or staging and as a tool to predict or monitor treatment response or toxicity. Human serum albumin (HSA) exists in both reduced and oxidized forms, and the percentage of oxidized albumin increases in several diseases. However, little is known regarding the pathological and physiological significance of oxidation due to poor characterization of the precise structural and functional properties of oxidized HSA. Here, we characterize both the structural and functional differences between reduced and oxidized HSA. Using LC-ESI TOF MS and FT MS analysis, we determined that the major structural change in oxidized HSA in healthy human plasma is a disulfide-bonded cysteine at the thiol of Cys34 of reduced HSA. Based on this structural information, we prepared standard samples of purified HSA, e.g. non-oxidized (intact purified HSA which mainly exists in reduced form), mildly oxidized and highly oxidized HSA. Using these standards, we demonstrated several differences in functional properties of HSA including protease susceptibility, ligand-binding affinity and antioxidant activity. From these observations, we conclude that an increased level of oxidized HSA may impair HSA function in a number of pathological conditions. In addition, we determined blood and plasma sampling conditions for the accurate measurement of the oxidized albumin ratio in plasma by using EST-TOF MS screening.
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藏滿 保宏, 岩本 早耶香, 田場 久美子, 良沢 昭銘, 藤本 正憲, 沖田 極, 坂井田 功, 中村 和行原稿種別: シンポジウム1：臨床プロテオミクスの現状と将来像―網羅的から選択的解析へ／定性から定量的解析へ―
2008 年 52 巻 2 号 p. 31-34
発行日: 2008年
公開日: 2009/12/04
ジャーナル フリーWhile Gemcitabine (2'-deoxy-2'-difluorodeoxycytidine: Gemzar) (GEM) is thought to be most effective for pancreatic cancer, its clinical effects do not seem properly evaluated due to a high level of inherent and acquired resistance in the tumor cells. We analyzed GEM-resistant and GEM-sensitive human pancreatic cancer cell lines proteomically to identify the proteins related to the GEM-resistance. First, we separated cancer cell proteins by two-dimensional gel electrophoresis, and identified the protein spots whose expressions differed between GEM-sensitive and -resistant cell lines by liquid chromatography-tandem mass spectrometry (LC-MS / MS). It was recognized that heat shock protein 27 (HSP27) was high in GEM-resistant cell lines, and that KLM1-R cells, a cell line which has acquired resistance to GEM, restored sensitivity to it when HSP27 was knocked down by siRNA. These findings suggest that expression of HSP27 should be reduced in order that the antitumor effects of GEM may be well exerted. Since the expression of HSP27 was dramatically reduced in the KLM1-R cells treated with GEM and IFN-γ, improved therapeutic effects may be expected in the combination of IFN-γ with GEM.
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中西 豊文, 田伏 洋子, 清水 章, 田窪 孝行原稿種別: シンポジウム1：臨床プロテオミクスの現状と将来像―網羅的から選択的解析へ/定性から定量的解析へ―
2008 年 52 巻 2 号 p. 35-38
発行日: 2008年
公開日: 2009/12/04
ジャーナル フリーTo identify cancer-associated antigens and their corresponding autoantibodies, we applied the novel approach, autoantibodiomics, established by Brichory et al., who reported annexins I and II as specific antigens in sera from adenocarcinoma patients. Solubilized proteins from cancer cell lines were separated using one or two dimensional polyacrylamide gel electrophoresis, followed by Western-blotting analysis, in which sera from individual patients were tested for primary antibodies. In cases of adenocarcinoma, we found several positive spots on PVDF membrane using a WB/enhanced chemiluminescence (ECLplus) detection kit, and identified 8 proteins, including α-enolase and peroxiredoxin IV in an adenocarcinoma A549 cell using matrix laser desorption time-of-flight mass spectrometry (MALDI-TOF MS). In cases of esophageal cancer, we also detected some positive spots on PVDF membrane and 2 proteins, peroxiredoxin-VI and heat shock 70 isoform in soluble proteins of a TE-2 cell by MALDI-TOF MS. Moreover, we detected non-Hodgkin’s lymphoma associated autoantibody in patients’ sera and identified L-plastin in the corresponding specific antigen derived from a Raji cell (one of B lymphoma cell lines). This technique, autoantibodiomics, was a powerful tool for the identification of candidate cancer-associated antigens and could be applied for the detection of these antigens and autoantibodies regardless of the selected cancer cell lines.
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