蛋白质芯片技术在矽肺早期血清生物标志物筛选中的应用

Screening of the serum biomarkers for early pneumoconiosis by SELDI-TOF-MS

目的寻找可对矽肺患者进行早期诊断的血清特异性生物标志物。方法选择接触矽尘未患病者(0组)、接触矽尘有可疑矽肺者(0+组)、接触矽尘后患I期矽肺者(I组)以及未接触矽尘的正常人(正常对照组)各20例,利用表面加强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测各组血清蛋白质指纹图谱。结果质荷比(M/Z)为2746、3269、3402、3941、4184、5488、5914、6122、7572、7937的10个标志分子在矽尘暴露人员(0、0+、I组)和正常人的血清中差异表达,相对含量变化具有统计学意义(P<0.05)。其中M/Z为2746的分子在矽尘暴露人员血清中高表达,而在正常人血清中低表达;相反,M/Z为4184、5914和6122的分子在正常人血清中高表达,而在矽尘暴露人员血清中低表达。M/Z为2938、3269、3821、4095、5071、5483、5640的7个标志分子在矽尘暴露人员各组血清中差异表达,相对含量变化具有统计学意义(P<0.05),其中M/Z为5071、5483和5640的分子在矽尘暴露人员0组和0+组中均为低表达,而在I组中高表达。结论M/Z为2746、4184、5914和6122的分子可能成为矽肺患者与正常人的鉴别诊断指标,而M/Z为5071、5483和5640的分子可能成为矽肺的早期诊断指标。

Objective To search the specific biomarkers in serum of patients with silicosis for early diagnosis. Methods Surface enhanced laser desorption ionization-time of flight-mass spectrometry （SELDI-TOF-MS） was used to search the specific biomarkers in serum samples from 60 silica-exposed workers （20 with no silicosis served as 0 group, 20 with suspected silicosis as 0＋ group and 20 with stage I silicosis as stage I group）, and 20 healthy men with no silica exposure as control. Serum samples were analyzed using the new generation of weak cationic exchange chip （CM10, Ciphergen Biosystems, America） and the protein fingerprint automatically collected using ProteinChip Biology System （PBSII-C） instrumentation. The data were analyzed by Ciphergen ProteinChip and BioMarker Wizard software. Results Compared with control group, ten biomarkers were differentially expressed among all the silica-exposed workers （P〈0.05）, including M/Z of 2 746, 3 269, 3 402, 3 941, 4 184, 5 488, 5 914, 6 122, 7 572 and 7 937. Compared with the control group, there existed a higher expression of protein with M/Z of 2 746 in 0, 0＋, I stage group. On the contrary, there was lower expression of proteins with M/Z of 4 184, 5 914 and 6 122 among silica-exposed groups compared with the control group. Interestingly, there were 7 different degrees of expression of proteins of M/Z 2 938, 3 269, 3 821, 4 095, 5 071, 5 483 and 5 640 among the silica-exposed workers （P〈0.05）. The levels of three molecular masses of 5 071, 5 483 and 5 640 were lower in 0 and 0＋ group than that of the I stage group. Conclusion Proteins with M/Z of 2 746, 4 184, 5 914 and 6 122 can be used to differentiate the patients with silicosis from healthy individuals based on present study, and proteins with M/Z of 5 071, 5 483 and 5 640 may be the potential biomarkers suggesting the diagnosis of early stage of silicosis.