MALDI-TOF-MS技术筛查原发性小肝细胞癌血清特异蛋白

目的应用基质辅助激光解析/电离飞行时间质谱(MALDI-TOF-MS)技术对原发性小肝细胞癌(小肝癌)患者与正常人血清蛋白质谱进行分析,得到高敏感性和特异性的血清蛋白,且能将小肝癌与正常人较准确的分开,建立诊断模型指纹图谱。方法采用德国Bruker公司的基质辅助激光解析电离质谱仪(MALDI-TOF-MS)分别检测35例原发性小肝癌患者与50名正常人血清蛋白质相对含量,运用FlexAnalysis和BioTools软件对两组血清蛋白质谱进行分析,寻找差异蛋白,再通过分析软件对其进行交叉验证,得到具有诊断意义的特异性蛋白标志物。结果检测到蛋白质质荷比(M/Z)在0～100000 Da范围内,小肝癌组与正常人组有差异的蛋白质峰为55个(P<0.05),交叉验证结果显示以M/Z值为7771.17 Da的蛋白质,具有较高敏感性和特异性,可将正常人及小肝癌患者准确分开,且M/Z值为7771.17 Da蛋白质在小肝癌组中低表达,在正常人组中高表达。再以M/Z值为7771.17Da的蛋白质作为生物标记,随机抽取小肝癌与正常人血清标本进行双盲验证,结果表明M/Z值为7771.17Da蛋白质作为生物标记物可以将小肝癌与正常人准确地分组,其敏感性为91.0%,特异性为70.0%。结论 MALDI-TOF-MS技术对小肝癌患者组、正常人组血清蛋白质谱进行分析,可得到原发性小肝癌早期诊断特异性、敏感性较高的血清特异性蛋白。

Application of MALDI-TOF-MS technique to screen the serum specific protein in SHCC

Objective By using the technique of matrix assisted laser desorption/ionization-time of flight-mass spectrometry(MALDI-TOF-MS) to analyse the serum protein spectrum and to get high sensitivity and specificity protein marker in SHCC patients and normal persons,then to establish serum markers pattern for the diagnosis of SHCC.Methods The technique of MALDI-TOF-MS made in Bruker company in German was used to detect relative content of serum protein in 35 cases of SHCC and 50 normal persons,and FlexAnalysis and BioTools software was used to analyse protein spectrum in two groups to find specific proteomic,then through the analysis software for cross-validation to get a diagnosis of specific proteins.Results There were 55 proteins peak having significant discrepancy in the 0-100000Da(P<0.05).The cross-validation results showed that the proteins(M/Z=7771.17Da) with high sensitivity and specificity could separate SHCC group from normal group accurately.The proteins(M/Z=7771.17Da) was lowly expressed in group of SHCC and highly expressed in group of normal persons.Then randomly selected SHCC and normal serum samples were taken to validate this protein(M/Z=7771.17Da) as biological markers could accurately separate SHCC proteins and normals.The sensitivity and specificity was 91.0% and 70.0%.Conclusion Using MALDI-TOF-MS to analyze serum protein spectrum in SHCC group and normal group could get the high specificity and sensitivity specific protein for early diagnosis of SHCC.