两个新的前列腺癌相关分泌蛋白的鉴定和表达.pdfVIP

HEREDITAS (Beijing) 2010 3 , 32(3): 235―241 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2010.00235 两个新的前列腺癌相关分泌蛋白的鉴定和表达 钱晓龙, 施庆国, 庞博, 武瑞琴, 俞岚, 李山虎, 王洪涛, 周建光 , 100850 为了获得代表不同前列腺癌进展阶段的细胞系的胞外蛋白表达谱, 验证其中差异表达蛋白是否为分泌 蛋白, 在细胞水平看其是否有作为前列腺癌血清标志物的潜质, 文章利用双向电泳寻找胞外蛋白中差异表达的 点, 并质谱鉴定其是何种蛋白质。应用RT-PCR 方法分析候选分子在8 种细胞系中的表达和对雄激素刺激的应 答, 构建了候选分子的真核表达载体, 瞬时转染293T 细胞, 应用标签抗体Western blotting 方法检测验证细胞培 养基中候选分子的表达。结果表明: 筛选出两个C4-2 胞外高表达的分子—— 磷酸丙糖异构酶-1(Triosephosphate isomerase 1, TPI1)和多配体聚糖结合蛋白(Syndecan binding protein, syntenin, ST1); 转录水平发现它们与前列腺 癌恶性程度相关, 并且后者受雄激素的作用下调; 二者均为分泌蛋白。磷酸丙糖异构酶-1 和多配体聚糖结合蛋 白均有作为指示前列腺癌发展阶段的血清标志物的潜质。 前列腺癌; 血清标志物; 分泌蛋白; 磷酸丙糖异构酶-1; 多配体聚糖结合蛋白 Identification and expression of two new secretory proteins associated with prostate cancer QIAN Xiao-Long, SHI Qing-Guo, PANG Bo, WU Rui-Qin, YU Lan, LI Shan-Hu, WANG Hong-Tao, ZHOU Jian-Guang The Laboratory of Molecular Biology of Medicine, Beijing Institute of Biotechnology, Beijing 100850, China Abstract: Our research intends to obtain extra-cellular proteinogram of cell lines representing different advancement stages of prostate cancer and to test whether screened differential expression proteins can be secreted and used as serum biomarkers for prostate cancer. By examining differential expression spots in two extra-cellular protein 2Dgels and mass spectrum, candidate molecules were obtained. The expressions of these candidate molecules in eight cell lines and response to androgen stimulus in LNCaP were analyzed by RT-PCR. By constructing eukaryotic expression vectors and western-blotting with anti tags antibodies, the candidate molecules were tested to understand whether they can be expressed in t