磁珠联合基质辅助激光解吸电离飞行时间质谱技术检测肾透明细胞癌尿液差异蛋白的初步应用

doi:10.24920/J1001-9294.2017.046

Chinese Medical Sciences Journal ›› 2017, Vol. 32 ›› Issue (4): 248-252.doi: 10.24920/J1001-9294.2017.046

磁珠联合基质辅助激光解吸电离飞行时间质谱技术检测肾透明细胞癌尿液差异蛋白的初步应用

董德鑫, 纪志刚^*(), 李汉忠, 严维刚, 张玉石

中国医学科学院北京协和医学院,北京协和医院泌尿外科,北京100730

收稿日期:2017-03-17 出版日期:2017-12-30 发布日期:2017-12-30
通讯作者: 纪志刚 E-mail:jzg1129@medmail.com.cn

Preliminary Application of WCX Magnetic Bead-Based Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry in Analyzing the Urine of Renal Clear Cell Carcinoma

Dong Dexin, Ji Zhigang^*(), Li Hanzhong, Yan Weigang, Zhang Yushi

Department of Urology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China

Received:2017-03-17 Published:2017-12-30 Online:2017-12-30
Contact: Ji Zhigang E-mail:jzg1129@medmail.com.cn

摘要/Abstract

摘要： 目的

弱阳离子磁珠联合基质辅助激光解吸电离飞行时间质谱( matrix-assisted laser desorption /ionization time of flight mass spectrometry,MALDI-TOF MS) 技术对肾透明细胞癌进行尿液差异蛋白研究,寻找尿液中特征性差异蛋白,评估该技术对肾透明细胞癌早期诊断的价值。

方法

收集活检证实的新发肾透明细胞癌患者11例（男10例,女1例,年龄46-78岁,平均63.0岁）,以及健康志愿者10例（均男性,年龄25-32岁,平均28.7岁）。采集病例组和对照组晨尿, 应用磁珠联合MALDI-TOF MS技术检测尿液中的蛋白质谱,利用ClinProTools2.2软件通过遗传算法和支持向量机确认两组尿液中的差异蛋白,筛选对肾透明细胞癌具有诊断价值的特征蛋白。

结果

肾透明细胞癌癌患者与健康组尿液内存在差异蛋白峰160个, 其中具有显著性差异的蛋白峰1个（2221.71 Da, P=0.0304）。遗传算法和支持向量机优化选择发现有13个差异蛋白峰对诊断预测模型有价值。

结论

磁珠联合MALDI-TOF MS技术检测尿液差异蛋白可能在肾透明细胞癌的早期诊断方面具有一定的临床应用价值。

关键词: 肾透明细胞癌, 蛋白质组学, 磁珠, 尿液

Abstract: Objective

To evaluate the application of weak cation exchange (WCX) magnetic bead-based Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) in detecting differentially expressed proteins in the urine of renal clear cell carcinoma (RCCC) and its value in the early diagnosis of RCCC.

Methods

Eleven newly diagnosed patients (10 males and 1 female, aged 46-78, mean 63 years) of renal clear cell carcinoma by biopsy and 10 healthy volunteers (all males, aged 25-32, mean 29.7 years) were enrolled in this study. Urine samples of the RCCC patients and healthy controls were collected in the morning. Weak cation exchange (WCX) bead-based MALDI-TOF MS technique was applied in detecting differential protein peaks in the urine of RCCC. ClinProTools2.2 software was utilized to determine the characteristic proteins in the urine of RCCC patients for the predictive model of RCCC.

Results

The technique identified 160 protein peaks in the urine that were different between RCCC patients and health controls; and among them, there was one peak (molecular weight of 2221.71 Da) with statistical significance (P=0.0304). With genetic algorithms and the support vector machine, we screened out 13 characteristic protein peaks for the predictive model.

Conclusions

The application of WCX magnetic bead-based MALDI-TOF MS in detecting differentially expressed proteins in urine may have potential value for the early diagnosis of RCCC.

Key words: renal clear cell carcinoma, proteomics, magnetic beads, urine, diagnosis model

Dong Dexin, Ji Zhigang, Li Hanzhong, Yan Weigang, Zhang Yushi. Preliminary Application of WCX Magnetic Bead-Based Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry in Analyzing the Urine of Renal Clear Cell Carcinoma[J].Chinese Medical Sciences Journal, 2017, 32(4): 248-252.

图/表 4

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Index	Molecular mass (Da)	Start mass (Da)	End mass (Da)	Weight in diagnostic model
45	2544.23	2539.39	2547.26	1.48
46	2549.69	2547.26	2554.68	1.11
77	3675.59	3665.07	3680.1	1.31
85	4010.08	4004.66	4020.37	0.24
92	4270.51	4262.22	4280.24	1.23
98	440.75	4417.21	4431.27	0.60
102	4820.53	4812.39	4830.27	0.89
128	6635.97	6621.02	6643.5	1.04
137	8015.2	8007.3	8022.17	0.10
147	8659.21	8646.72	8666.43	1.33
150	9013.4	9005.82	9019.82	0.83
154	9198.44	9190.34	9204.48	0.31
160	9842.68	9833.65	9825.85	0.33

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磁珠联合基质辅助激光解吸电离飞行时间质谱技术检测肾透明细胞癌尿液差异蛋白的初步应用

Preliminary Application of WCX Magnetic Bead-Based Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry in Analyzing the Urine of Renal Clear Cell Carcinoma

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