Then analysis of lung cancer protein using two-dimensional polyacrylamide gel electrophoresis Application for adjuvant chemotherapy

二维聚丙烯酰胺凝胶电泳分析肺癌蛋白在辅助化疗中的应用

• 批准号: 10671271
• 负责人: HIRANO Takashi
• 金额: $ 1.92万
• 依托单位: Tokyo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671271/
• 关键词: lung cancer protein; antibody; prognosis; TA02; NapsinA; reticulocalbin; GST-π; Two-dimensional polyacrylamide gel electrophoresis; 手術予後; reticulocarbin; 原発性肺癌

1. Sixteen polypeptides associated with histopathological differentiation were detected using two-dimensional polyacrylamide gel electrophoresis of primary lung carcinoma.2. Based on the expression of the polypeptides associated with either histopathological differentiation or cell proliferation, we classified primar lung adenocarcinoma into typical case and atypical case. 65.4% of the typical cases were diagnosed as well differentiated adenocarcinoma histopathologically. Also, 65.4% of the typical cases showed negative in lymph node metastasis. On the other hand, 20.5% and 34.9% of the atypical cases were judged as well differentiated case and cases without lymph node metastasis, respectively. Furthermore, we recognized the statistically significant difference in three year-disease free survival(62.9% in typical cases, 27.1% in atypical cases ; p=0.0011).3. We produced monoclonal antibodies against N-terminal of TA02 molecule. The histological distribution of TA02 was clarified immuno … More histochemically using this antibody and human tissues. Also, we obtained the evidence that TA02 molecule was homologos with NapsinA which was a novel aspartic proteinase published at the end of 1998, because the adenocarcinoma cases with high expression of TA02 showed the expression of napsin A in RNA level. Furthermore, GST fusion protein of Napsin A showed positive reaction with anti-TA02 antibody.4. TS02 associated with neuroendocrine cells was homologous with protein gene product 9.5(PGP 9.5). In the TS02 positive cases two-years prognosis become worse compared to TS02 negative cases.5. Compred to the 2DE patterns between CDDP-resistant cell line and the parents cell line, the positions of reticulocarbin and glutathione-S-transferase π were identified on 2DE gel. The expession level of reticulocalbin, which was Ca-binding protein at endoplasmic reticulum, became low in the CDDP-resistant cell line. On the other hand, the expression level of GST-πbecame high, remarkably.6. The analysis based on lung cancer protein using 2DE is valuable for the evaluation of biological tumor malignancy. Less

1.采用双向聚丙烯酰胺凝胶电泳检测16种与肺癌组织分化相关的多肽.根据与组织病理学分化或细胞增殖相关的多肽的表达，我们将原发性肺腺癌分为典型病例和不典型病例。65.4%的典型病例组织病理学诊断为高分化腺癌。65.4%的典型病例淋巴结转移阴性。另一方面，20.5%和34.9%的不典型病例分别被判定为高分化病例和无淋巴结转移病例。此外，我们认识到三年无病生存率的统计学显著差异（典型病例为62.9%，非典型病例为27.1%; p=0.0011）。我们制备了针对TA 02分子N端的单克隆抗体。用免疫组化方法明确了TA 02的组织学分布。 ...更多信息 使用这种抗体和人体组织进行组织化学分析。TA 02高表达的腺癌病例在RNA水平上也有napsin A的表达，这也证明了TA 02与1998年底发表的一种新的天冬氨酸蛋白酶NapsinA具有同源性。Napsin A的GST融合蛋白与抗TA 02抗体呈阳性反应.与神经内分泌细胞相关的TS 02与蛋白质基因产物9.5（PGP 9.5）同源。TS 02阳性病例的2年预后较阴性病例差.通过比较CDDP抗性细胞系与亲本细胞系之间的双向电泳图谱，确定了网织红蛋白和谷胱甘肽-S-转移酶π在2DE上的位置。CDDP耐药细胞内质网钙结合蛋白网织红蛋白表达水平降低。另一方面，GST-π的表达水平明显升高.基于肺癌蛋白质的二维凝胶电泳分析对生物学肿瘤恶性程度的评价具有重要价值。少