Bladder cancer associated glycoprotein signatures revealed by urinary proteomic profiling
Bladder cancer associated glycoprotein signatures revealed by urinary proteomic profiling
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DOI：
10.1021/pr0700807
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发表时间：
2007-01-01
影响因子：
4.4
通讯作者：
Goodison, Steve
中科院分区：
文献类型：
--
作者：
Kreunin, Paweena;Zhao, Jia;Goodison, Steve
目前通过尿液分析无创检测和监测膀胱癌的方法包括：空尿细胞学（VUC）和一些诊断性尿蛋白生物标志物；然而，由于VUC的敏感性较差和目前可用的蛋白质检测的高假阳性率，通过尿液分析检测膀胱癌仍然是一个挑战。在这里提出的研究中，开发了一种快速，高灵敏度的技术来分析自然微量人类尿液标本中的n -连接糖蛋白成分。在进行线性离子阱质谱分析之前，利用刀豆蛋白A （cona）亲和层析耦合纳米流液相层析分离复合肽混合物。通过多次分析鉴定出的186种高可信度蛋白中，40%为分泌蛋白，18%为膜蛋白，14%为细胞外蛋白。在这项研究中，几种蛋白质的存在似乎与膀胱癌的存在有关，包括在所有荷瘤患者样本中检测到的α - 1b糖蛋白，但在从非荷瘤个体获得的样本中均未检测到。Con A亲和层析和纳米lc /MS/MS的结合提供了复杂生物样品中n -糖蛋白的初步研究，并有助于在无创获取的人类尿液中鉴定潜在的膀胱癌生物标志物。
Current methods in the noninvasive detection and surveillance of bladder cancer via urine analysis include voided urine cytology (VUC) and some diagnostic urinary protein biomarkers; however, due to the poor sensitivity of VUC and high false-positive rates of currently available protein assays, detection of bladder cancer via urinalysis remains a challenge. In the study presented here, a rapid, high-sensitivity technique was developed to profile the N-linked glycoprotein component in naturally micturated human urine specimens. Concanavalin A (Con A) affinity chromatography coupled to nanoflow liquid chromatography was utilized to separate the complex peptide mixture prior to a linear ion trap MS analysis. Of 186 proteins identified with high confidence by multiple analyses, 40% were secreted proteins, 18% membrane proteins, and 14% extracellular proteins. In this study, the presence of several proteins appeared to be associated with the presence of bladder cancer, including alpha-1B-glycoprotein that was detected in all tumor-bearing patient samples but in none of the samples obtained from non-tumor-bearing individuals. The combination of Con A affinity chromatography and nano-LC/MS/MS provides an initial investigation of N-glycoproteins in complex biological samples and facilitates the identification of potential biomarkers of bladder cancer in noninvasively obtained human urine.