Exhaustive proteome analyses of pulmonary neuroendocrine carcinomas

肺神经内分泌癌的详尽蛋白质组分析

基本信息

批准号：
19590365
负责人：
SATO Yuichi
金额：
$ 2.91万
依托单位：
Kitasato University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2007
资助国家：
日本
起止时间：
2007 至 2009
项目状态：
已结题

项目摘要

I describe recent results of the search for pulmonary neuroendocrine carcinoma markers with diverse proteome techniques.1. Exhaustive monoclonal antibodies production: We generated monoclonal antibodies using cells derived from pulmonary carcinomas as an immunogen. From a group of obtained antibodies, present described antibody for the receptor of activated C kinase 1 (RACK1). The expression RACK1 was significantly high and frequent in adenocarcinomas but was barely detected in a few squamous cell carcinomas and large cell carcinomas. Moreover, RACK1 expression was also significantly associated with the pathological stage, tumor size and lymph node status of adenocarcinoma patients. These results suggest that RACK1 may be a novel differential diagnostic marker for pulmonary adenocarcinomas. 2. Detection of peptides in the conditioned medium from carcinoma cell lines: Aiming to identify new markers of pulmonary neuroendocrine tumors, we analyzed comprehensively analyzed peptides which w … More ere secreted into the conditioned medium by LCN1, an LCNEC line. Two peptide fragments of 40 and 19 amino acid residues were identified by MALDI-TOF/TOF MS. These two fragments were demonstrated to be parts of VGF. RT-PCR analysis of lung cancer cell lines showed that vgf was expressed only in neuroendocrine carcinoma-derived cells. Our data suggest that VGF can be a novel sero-diagnostic marker of pulmonary neuroendocrine tumors. 3. Application of two-dimensional gel electrophoresis: We studied the proteomic approach using cell lysate from two cell lines (N231 derived from small cell lung carcinoma (SCLC) and LCN1 derived from pulmonary large cell neuroendocrine carcinoma (LCNEC)) with two-dimensional gel electrophoresis (2-DE) and MULDI-TOF/TOF MS. Within the 25 identified proteins, cytokeratins (CK) 7, 8, 18 and 19 were up-regulated in LCN1 cells compared with N231 cells. Their expressions were studied immunohistochemically with 81 pulmonary carcinomas. The mean immunostaining scores of CK7, 8, 18, and 19 were significantly higher in LCNEC cases than in SCLC. These data suggest that the biological characteristics of LCNEC and SCLC may be different and the expressions of CKs may serve as differential diagnostic markers. 4. Detection of autoantibodies in sera of patients with lung cancer: To detect autoantibodies (AAs), sera from SCLC patient was screened by immunoblotting using cell lysate of four cell lines. To identify the antigens recognized by AAs, two-dimensional gel electrophoresis was immunoblotted and target spots were cut out from the membrane and gel. By this method, villin1 was identified with AA in sera from patients with SCLC. Villin1 may be useful markers to distinguish LCNEC/AD from SCLC/SCC, and the present method might be useful to identify specific tumor-associated molecules in sera from pulmonary carcinoma patients with different histologic types. Less

我描述了具有多种蛋白质组技术的肺神经内分泌标记的结果：我们使用肺癌的细胞产生了单克隆抗体RACK1在腺癌中显着且频繁，但在少数鳞状癌中几乎没有检测到，RACK1表达也与病理学阶段，肿瘤IZE和淋巴结症患者的腺癌患者可能相关。诊断标志物folmon Rcinomas。通过MALDI-TOF/TOF，这两个片段被证明是仅在神经内分泌癌衍生的细胞中的肺癌细胞的VGF RT-PCR分析的一部分。蛋白质组学方法使用细胞系（N231衍生自小肺癌（SCLC）衍生1衍生的1 ry大细胞神经内分泌癌（LCNEC），具有二维凝胶电泳（2-DE）和Muldi-TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF/TOF。 S> </s> </s> </s> </s> </s> </s> </s>离子通过81个肺癌的免疫组织化学研究了。 LCNEC Casses比SCLC中的数据表明，LCNEC和SCLC的生物学特征可能是较高的，CKS的表达可能是差异诊断标记。 ES。鉴定AAS识别的抗原，通过这种方法从膜和凝胶中切出了两种身份的凝胶电泳，而靶斑是SCLC患者的ITH AA SCLC/SCC，目前的方法可能有助于鉴定具有不同组织学类型的肺癌患者的血清中特定的肿瘤相关分子。