应用液体芯片-质谱技术筛选肺鳞癌患者血清标志蛋白

Application of liquid chip-mass spectrometry technology for screening serum biomarker proteins in lung squamous cell carcinoma

目的探讨液体芯片．质谱技术（CLINPROT系统）在肺鳞癌患者血清标志蛋白筛选中的应用。方法应用CLINPROT系统检测34例肺鳞癌患者（肺鳞癌组）、46例良性肺疾病患者（良性对照组）及44名正常人（正常对照组）的血清蛋白表达谱，应用FlexAnalysis3．0软件进行数据分析，筛选差异表达蛋白质；采用液相色谱串联质谱（LC—MS／MS）技术鉴定标志蛋白。结果分别比较肺鳞癌组与正常对照组以及肺鳞癌组与良性对照组血清蛋白表达谱，在质荷比（M／Z）800—10000范围内，前者筛选出96个差异表达蛋白峰，其中M／Z为4054．13和4267．46的蛋白峰在2组间差异最大，用这2种差异蛋白建立的坐标系具有良好的区分肺鳞癌与正常人的能力；后者筛选出99个差异表达蛋白峰，其中M／Z为5065．27和4054．02的蛋白峰在2组间差异最大，用这2种差异蛋白建立的坐标系具有良好的区分肺鳞癌与良性肺疾病的能力。取M／Z为1778和1865的蛋白行LC-MS／MS鉴定，结果提示二者可能均为补体C3片段或C3前体。结论肺鳞癌患者与正常人以及与良性肺疾病患者之间血清蛋白表达谱存在明显差异，应用液体芯片-质谱技术可能从血清中筛选出敏感性高、特异性强的肺鳞癌标志蛋白。

Objective To screen the serum biomarker proteins of lung squamous cell carcinoma （SCCs） by liquid chip-mass spectrometry technology. Methods All serum samples, including 34 SCCs, 46 benign lung diseases （BLDs） and 44 healthy individuals, were analyzed by CLINPROT system in order to study the serum protein expression profiles. Then the discriminatory proteins were detected by FlexAnalysis 3.0 software. Biomarkers were identified by liquid chromatography-tandem mass spectrometry （LC- MS/MS）. Results Comparing the differential serum expression proteins between SCCs and healthy individuals, and SCCs and BLDs respectively. Ninty-six differential protein peaks [ mass-to-charge ration （M/Z） between 800 and 10 000] were found between SCCs and healthy individuals. In these protein peaks, the expression of protein peaks at 4054. 13 M/Z and 4267.46 M/Z had the largest difference between them. The two protein peaks could accurately separate SCCs from healthy individuals by the frame of axes. Similarly, 99 differential protein peaks were automatically detected between SCCs and BLDs. In these protein peaks, the expression of protein peaks at 5065.27 M/Z and 4054.02 M/Z had the largest difference between them. The two protein peaks could accurately separate SCCs from BLDs by the frame of axes. Identified by LC-MS/MS, 1778 M/Z and 1865 M/Z might be assayed jointly and corresponded to complements C3 fragment or C3f precursor. Conclusions Differential protein expressions existed between SCCs versus healthy individuals and SCCs versus BLD patients. It is feasible to screen the diagnostic serum biomarkers of SCC with a high sensitivity and specificity by using CLINPROT system.