Biomarkers for Prostate and Bladder Cancer

前列腺癌和膀胱癌的生物标志物

• 批准号: 6951555
• 负责人: ALVIN Y LIU
• 金额: $ 45.9万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-22 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6951555
• 关键词: biomarker; bladder neoplasm; clinical research; cooperative study; diagnosis design /evaluation; glycopeptides; human subject; in situ hybridization; mass spectrometry; neoplasm /cancer diagnosis; oncoproteins; patient oriented research; prostate neoplasms; proteomics; urinalysis; urine; western blottings

DESCRIPTION (provided by applicant): A strategy based on mass spectrometry proteomics is used to discover biomarkers for prostate and bladder cancer. The markers targeted are proteins secreted by the cancer cells and/or cancer-associated cells. Tumor tissue specimens are obtained from surgically resected organs after pathology. The specimens are digested by collagenase into single cells in a serum-free media. The digestion media, termed COL, is made cell-free by centrifugation, and it contains proteins made by the cells. A matched non-cancer specimen is processed similarly for comparative analysis. Differentially expressed protein species in the cancer and non-cancer COL pairs are screened by a method termed glycopeptide capture followed by tandem mass spectrometry for identification. Since virtually all secreted proteins are post-translationally modified by the addition of carbohydrate groups this method selectively analyzes these proteins. Quantification is achieved by isotopic labeling of the peptides, i.e. light isotopes for cancer and heavy isotopes for non-cancer. Candidate markers will be validated by Western blotting of the COL samples and immunohistochemistry if specific antibodies are available, or by in situ nucleic acid probe hybridization. This marker discovery process is illustrated by TIMP1, found in our initial experiments. The feasibility of proteomic analysis of urine samples to detect markers will be done. Preliminary experiments have generated a human urinary proteome of 239 protein species. Urine samples will be obtained from cancer patients and age-matched volunteers. The goal is to develop an assay for cancer associated protein markers that can be detected in voided urine for the diagnosis of prostate and bladder cancer.

描述(申请人提供)：一种基于质谱学蛋白质组学的策略被用于发现前列腺癌和膀胱癌的生物标记物。目标标记物是癌细胞和/或癌症相关细胞分泌的蛋白质。肿瘤组织标本取自病理后手术切除的器官。胶原酶将标本在无血清培养基中消化成单细胞。消化介质，称为Col，通过离心法使其不含细胞，它含有由细胞产生的蛋白质。对匹配的非癌症样本进行类似的处理以进行比较分析。通过一种称为糖肽捕获的方法筛选癌症和非癌症Col对中差异表达的蛋白质种类，然后进行串联质谱分析以进行鉴定。由于几乎所有分泌的蛋白质都是通过添加碳水化合物基团进行翻译后修饰的，这种方法选择性地分析了这些蛋白质。定量是通过多肽的同位素标记实现的，即癌症的轻同位素和非癌症的重同位素。候选标记将通过Col样本的Western blotting和免疫组织化学(如果有特定抗体可用)或通过原位核酸探针杂交来验证。TIMP1说明了这个标记发现过程，它是在我们最初的实验中发现的。将对尿样进行蛋白质组学分析以检测标志物的可行性。初步实验已经产生了一个包含239种蛋白质的人类尿液蛋白质组。尿样将从癌症患者和年龄匹配的志愿者那里获得。其目标是开发一种可以在排尿中检测到癌症相关蛋白标记物的分析方法，用于诊断前列腺癌和膀胱癌。