Isolation and identification of potential urinary microparticle biomarkers of bladder cancer
Isolation and identification of potential urinary microparticle biomarkers of bladder cancer
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DOI：
10.1021/pr700775x
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发表时间：
2008-05-01
影响因子：
4.4
通讯作者：
Theodorescu, Dan
中科院分区：
文献类型：
--
作者：
Slrnalley, David M.;Sheman, Nicholas E.;Theodorescu, Dan
膀胱癌在美国每年导致大约13000人死亡。早期疾病通常以最低的发病率进行治疗，预后良好，而晚期疾病则相反。目前，还没有早期发现这种癌症的工具。微粒是小的亚细胞颗粒，基本上由所有细胞在活化时释放，并且已知由癌细胞组成性产生。由于大多数膀胱癌起源于器官内腔的尿路上皮细胞，我们假设这些细胞将释放微粒到尿液中。本研究的目的是确定膀胱癌患者尿液微粒中的潜在生物标志物。从5个健康个体和4个膀胱癌个体中分离尿微粒。通过PAGE和胰蛋白酶消化对样品进行脱脂，提取肽，并使用Thermo Finnigan LTQ和LTQ-FT离子阱质谱仪通过LC-MS/MS检查蛋白质组。通过SEQUEST确定蛋白质鉴定，并通过比较光谱计数评估相对定量。膀胱癌患者的微粒中有8种蛋白质升高。它们包括与表皮生长因子受体途径相关的五种蛋白质、GsGTP结合蛋白的α亚基、α-谷氨酰胺和视黄酸诱导蛋白3。需要进一步的研究来验证这些潜在的生物标志物。
Bladder cancer leads to approximately 13 000 deaths annually in the United States. Early disease is often treated with minimal morbidity and has good prognosis, while the opposite is true for advanced disease. Currently, no tools exist for early detection of this cancer. Microparticles are small, subcellular particles released by essentially all cells upon activation and are known to be produced constitutively by cancer cells. Since most bladder cancers originate in the urothelial cells lining the lumen of the organ, we hypothesize that these cells will release microparticles into the urine. The goal of this study was to identify potential biomarkers in the urinary microparticles of individuals with bladder cancer. Urine microparticles from five healthy individuals and four individuals with bladder cancer were isolated. Samples were delipidated by PAGE and trypsin-digested, peptides were extracted, and the proteome was examined by LC-MS/MS using a Thermo Finnigan LTQ and LTQ-FT ion trap mass spectrometer. Protein identification was determined by SEQUEST and relative quantitation was assessed by comparing spectral counts. Eight proteins were elevated in the microparticles from individuals with bladder cancer. They include five proteins associated with the epidermal growth factor receptor pathway, the alpha subunit of GsGTP binding protein, resistin, and retinoic acid-induced protein 3. Further studies will be needed to validate these potential biomarkers.