Department of Clinical Laboratory, Beijing Shijitan Hospital, Capital Medical University, China.
Clin Genitourin Cancer. 2013 Mar;11(1):56-62. doi: 10.1016/j.clgc.2012.06.003. Epub 2012 Sep 14.
We searched for bladder tumor markers by analyzing urine samples from patients with bladder cancer and from normal controls.
Proteins in urine samples of patients with bladder cancer and with normal controls were systematically examined by 2-dimensional electrophoresis combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The expression of the protein apolipoprotein A-I (apoA-I) was confirmed by Western blot analysis and further evaluated.
We successfully obtained the 2-dimensional electrophoresis gel maps of urinary proteins in patients with bladder cancer and in normal controls. Thirty differentially expressed protein spots were successfully matched by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Combined with the SWISS-PROT database, only 14 proteins (beta-2-microglobulin, fatty acid-binding protein adipocyte, gelsolin, isoform 1 of gelsolin, myoglobin, isoform 2 of fibrinogen alpha chain, apoA-I, prostaglandin D(2) synthase 21 kDa [brain], protein AMBP, transthyretin, keratin type II cytoskeletal 1, type II cytoskeletal 8, putative uncharacterized protein ALB, putative uncharacterized protein MASP2 [fragment]) were identified, including 2 putative proteins. Furthermore, apoA-I was confirmed by Western blot analysis, and the high level of apoA-I was found in urine samples from patients with bladder tumors compared with normal controls.
Analysis of urinary proteome may be a feasible, noninvasive, and efficient strategy for searching for potential bladder tumor biomarkers. A significant relationship of expressed apoA-I was established between bladder cancer and normal controls. We concluded that 14 differential spots included the apoA-I and would be potential urinary biomarkers for the diagnosis and surveillance of bladder cancer.
通过分析膀胱癌患者和正常对照者的尿液标本，寻找膀胱肿瘤标志物。
采用二维电泳结合基质辅助激光解吸电离飞行时间质谱技术系统检测膀胱癌患者和正常对照者尿液标本中的蛋白质，用 Western blot 分析进一步验证和评价其中载脂蛋白 A-I（apoA-I）的表达。
成功获得膀胱癌患者和正常对照者尿液蛋白质的二维电泳凝胶图谱，经基质辅助激光解吸电离飞行时间质谱成功匹配 30 个差异表达的蛋白质斑点，结合 SWISS-PROT 数据库，仅鉴定出 14 种蛋白质（β-2-微球蛋白、脂肪结合蛋白脂肪细胞、凝溶胶、凝溶胶同工型 1、肌红蛋白、纤维蛋白原α链同工型 2、apoA-I、前列腺素 D2 合酶 21kDa[脑]、蛋白 AMBP、转甲状腺素蛋白、角蛋白 II 细胞骨架 1、细胞骨架 8、假定未知蛋白 ALB、假定未知蛋白 MASP2[片段]），其中包括 2 种假定未知蛋白。进一步用 Western blot 分析证实了 apoA-I 的表达，膀胱癌患者尿液标本中 apoA-I 的水平明显高于正常对照者。
尿液蛋白质组分析可能是一种可行的、非侵入性的、有效的寻找潜在膀胱肿瘤标志物的策略。apoA-I 在膀胱癌和正常对照者之间存在显著的表达差异。我们推断，14 个差异斑点包括 apoA-I，可能成为膀胱癌诊断和监测的潜在尿液生物标志物。
