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尿蛋白质组学在膀胱尿路上皮癌早期诊断中的应用

[Application of urinary proteomics in early diagnosis of bladder urothelial carcinoma].

作者信息

Li Hong-jie, Li Chang-ying, Zhang Ting, Chen Juan-juan, Wang Shi-xin, Chang Ji-wu, Jiang Shou-fang, Zhang Guang-ling, Li Jian-Min

机构信息

Department of Pathophysiology, HeBei United University, Tangshan 063000, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2013 Apr;31(4):262-5.

Abstract

OBJECTIVE

To investigate the difference in urinary proteome between patients with bladder urothelial carcinoma (BUC) and healthy volunteers and to provide a basis for the early diagnosis of BUC.

METHODS

The urine samples from BUC patients and healthy volunteers (controls) were treated by 25% ethanol precipitation and two-dimensional gel electrophoresis (2-DE), and the obtained urinary proteins were subjected to Coomassie brilliant blue staining and analysis by PDQuest 8.0 (2-DE image analysis software); the differentially expressed proteins were sequenced by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry and identified using the Swiss-Prot database; the differential expression of these proteins was verified by western blot.

RESULTS

High-resolution and high-reproducibility 2-DE images were obtained from the urine samples of BUC patients and controls, with 789 ± 18 and 762 ± 14 protein spots, respectively. Compared with the control group, the BUC grouP had significantly decreased expression of 6 protein spots and significantly increased expression of 11 protein spots. The mass spectrometry revealed five proteins with increased expression in the BUC group, including fibrinogen, lactate dehydrogenase B, apolipoprotein A1, clusterin, and haptoglobin, and the results were confirmed by western blot.

CONCLUSION

There is significant difference in urinary proteome between BUC patients and healthy volunteers; the identification of differentially expressed proteins in urine lays the foundation for identifying potential molecular markers in early diagnosis of BUC.

摘要

目的

探讨膀胱尿路上皮癌(BUC)患者与健康志愿者尿液蛋白质组的差异,为BUC的早期诊断提供依据。

方法

采用25%乙醇沉淀法和二维凝胶电泳(2-DE)处理BUC患者和健康志愿者(对照组)的尿液样本,所得尿液蛋白质经考马斯亮蓝染色后,使用PDQuest 8.0(2-DE图像分析软件)进行分析;通过基质辅助激光解吸/电离飞行时间/飞行时间质谱对差异表达蛋白进行测序,并使用Swiss-Prot数据库进行鉴定;通过蛋白质印迹法验证这些蛋白的差异表达。

结果

从BUC患者和对照组的尿液样本中获得了高分辨率和高重现性的2-DE图像,分别有789±18和762±14个蛋白点。与对照组相比,BUC组有6个蛋白点表达显著降低,11个蛋白点表达显著升高。质谱分析显示BUC组有5种蛋白表达增加,包括纤维蛋白原、乳酸脱氢酶B、载脂蛋白A1、簇集素和触珠蛋白,蛋白质印迹法证实了该结果。

结论

BUC患者与健康志愿者的尿液蛋白质组存在显著差异;尿液中差异表达蛋白的鉴定为BUC早期诊断中潜在分子标志物的识别奠定了基础。

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