基于蛋白质组学的胸腔积液蛋白标志物筛选与验证

Proteomic techniques based protein biomarkers searching and validation in pleural effusion

目的用蛋白质组学技术对比分析良、恶性胸腔积液标本,寻找蛋白标志物为其鉴别诊断提供帮助和新线索。方法采用双向电泳分离、搜寻蛋白,基质辅助激光解吸飞行时间质谱(MALDI-TOF-MS)鉴定蛋白,ELISA验证蛋白在良恶性胸腔积液样本中具体含量。结果恶性组胶图对比良性组共显示明显差异蛋白点(上调或下调≥2倍)43个,上调9个,下调34个;对其中7个显著差异点(上调或下调≥3倍)质谱鉴定明确了具体类型;挑选显著差异点中免疫球蛋白λ(Igλ)、结合珠蛋白(Hp)进行ELISA验证,结果表明Igλ在良、恶性胸腔积液中含量差异无统计学意义,Hp含量组间差异有统计学意义(P<0.05)。进一步评估显示诊断标准为胸腔积液中Hp<389.02μg/L时,诊断恶性胸腔积液灵敏度为75.00%,特异度为52.38%。结论蛋白质组学技术的应用对胸腔积液蛋白标志物搜寻具有较大帮助,本研究搜寻到的标志物Hp对于良、恶性胸腔积液鉴别诊断具有一定价值,值得进一步研究。

Objective Through analyzing benign and malignant pleural effusion samples by proteomic techniques, finding protein biomarkers to provide help and new clues for effusion differential diagnosis. Methods Two-dimen- sional electrophoresis （2-DE） and matrix-assisted laser desorption ionization-time of flight-mass spectrometry （ MAL- DI-TOF-MS） were used to search and identify protein biomarkers, enzyme linked immunosorbent assay （ELISA） was to validate the biomarkers. Results By comparing malignant group with benign group, 43 significantly different protein spots（Up or down regulated≥2 times） were found. Including 9 up regulated spots and 34 down regulated spots. And 7 spots were identified（ Up or down regulated ≥3 times） by MALDI-TOF-MS and validated 2 spots im- munoglobulin λ（Igλ） and haptoglobin（Hp） by ELISA. The results showed that Igλ showed no statistical signifi- cance between two groups, while Hp showed the statistical significance（P 〈 0.05）. The diagnostic sensitivity and specificity of Hp in malignant pleural effusion were 75.00% and 52. 38% at diagnostic cut-off point of 389.02 μg/ L. Conclusion The application of proteomics technology has a great help with protein biomarkers searching in pleural effusion. HP has a certain value for the differential diagnosis of benign and malignant pleural effusionand and worthy of further study.