蛋白组技术发现和验证耐多药肺结核潜在血清标志物FCN3

Discovery and validation of potential biomarker FCN3 of multi-drug resistant pulmonary tuberculosis by protein group technology

目的利用血清蛋白组学技术筛选和鉴定耐多药肺结核患者血清差异表达蛋白。方法采用i TRAQ标记结合MALDI-TOF/MS筛选健康体检者、肺炎患者和耐多药肺结核患者血清的差异表达蛋白,并进行生物信息学分析;对差异蛋白FCN3进行MRM相对定量验证。结果一共筛选和鉴定到248种蛋白质,其中置信度在95%以上有92种;以正常对照者和肺炎患者血清为对照,在耐多药肺结核患者血清中发现和鉴定出29种差异表达蛋白(P<0.05),其中16种表达显著增高,13种表达显著降低,FCN3在耐多药肺结核患者血清中表达降低(P<0.05);MRM相对定量验证显示,与正常对照组相比,FCN3在肺炎和耐多药肺结核组中积分峰面积分别为0.818 5±0.037 7和0.378 6±0.020 2,FCN3在耐多药肺结核组表达水平明显降低(P<0.005)。结论 MRM验证FCN3的表达水平与血清蛋白组学筛选的结果一致,FCN3可能是耐多药肺结核潜在的血清标志物。

Objective Using serum proteomics technology to screen and identify the differential expression protein in patients with multidrug resistant pulmonary（ MDR- TB）. Methods Serum differentially expressed proteins from healthy objects,patients with MDR- TB and pneumonia patients were detected by i TRAQ labeling combined with matrix assisted laser desorption ionization time of flight mass spectrometry（ MALDI- TOF / MS） for further bioinformatical analysis. Results 248 proteins were dectected and identified,among which 92 proteins had the confidential levels above 95%. 29 differentially expressed proteins were detected and identified in the serum of patients with MDR- TB,comparing normal controls with patients with pneumonia（ P 0. 05）. The expression levels of 16 proteins significantly increased,and 13 proteins decreased in MDR- TB serum. The levels of FCN3 was reduced in the serum of patients with MDR- TB（ P 0. 05）. The result of MRM relative quantitiative verification showed that the integral peak area of FCN3 in pneumonia and MDR- TB were 0. 8185 ± 0. 0377 and 0. 3786 ± 0. 0202 respectively,when compared with the normal subjects. The expression level of FCN3 of MDR- TB significantly decreased in MDR- TB（ P 0. 01）. Conclusion The FCN3 level of MRM verification is consistent with serum proteomics screening.FCN3 might be a potential biomarker of MDR- TB.