Biomarkers for Prostate and Bladder Cancer

前列腺癌和膀胱癌的生物标志物

• 批准号: 7479816
• 负责人: ALVIN Y LIU
• 金额: $ 35.22万
• 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 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7479816
• 关键词: Age; Antibodies; Biological Assay; Biological Markers; Cancer Patient; Carbohydrates; Cells; Centrifugation; Diagnosis; Digestion; Glycopeptides; Goals; Human; Immunohistochemistry; In Situ; Isotopes; Label; Light; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Marker Discovery; Mass Spectrum Analysis; Methods; Nucleic Acid Probes; Organ; Pathology; Peptides; Process; Prostate; Proteins; Proteome; Proteomics; Resected; Sampling; Serum-Free Culture Media; Specimen; TIMP1 gene; Tumor Tissue; Urination; Urine; Western Blotting; base; cancer cell; collagenase; comparative; research study; tandem mass spectrometry; urinary; volunteer

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（由申请人提供）：基于质谱蛋白质组学的策略用于发现前列腺癌和膀胱癌的生物标志物。 靶向标记物是癌细胞和/或癌症相关细胞分泌的蛋白质。 肿瘤组织标本是在病理学后从手术切除的器官获得的。 标本在无血清培养基中被胶原酶消化成单细胞。 消化介质称为 COL，通过离心制成无细胞，并且含有细胞产生的蛋白质。 对匹配的非癌症样本进行类似处理以进行比较分析。 通过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，然后通过串联质谱进行鉴定。 由于几乎所有分泌的蛋白质都通过添加碳水化合物基团进行翻译后修饰，因此该方法选择性地分析这些蛋白质。 通过肽的同位素标记来实现定量，即用于癌症的轻同位素和用于非癌症的重同位素。 如果有特异性抗体，候选标记将通过 COL 样品的蛋白质印迹和免疫组织化学进行验证，或通过原位核酸探针杂交进行验证。 在我们最初的实验中发现的 TIMP1 说明了这个标记发现过程。 将进行尿液样本蛋白质组分析以检测标记物的可行性。 初步实验已经生成了包含 239 种蛋白质的人类尿液蛋白质组。尿液样本将从癌症患者和年龄匹配的志愿者中获取。 目标是开发一种癌症相关蛋白标记物的检测方法，可以在排泄的尿液中检测到该标记物，以诊断前列腺癌和膀胱癌。