应用蛋白质组学技术筛检肝癌血清差异蛋白质并分离鉴定阿朴脂蛋白AⅡ

目的 应用蛋白质组学技术筛选并鉴定肝癌患者血清中的差异蛋白质.方法 采用表面增强激光解析及电离飞行时间质谱技术,对33例肝癌患者和33例正常对照的血清蛋白质进行了检测分析,筛选肝癌的差异蛋白;用等电点沉淀法、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳及高效液相色谱串联质谱等一系列方法分离鉴定差异蛋白质.肝癌组与正常组蛋白质峰差异比较采用两样本t检验. 结果 肝癌组与正常人组血清蛋白质图谱在相对分子质量为2 000～10 000范围内,有65个蛋白质峰差异有统计学意义(P<0.05),由此组建的诊断模型灵敏度为100%,特异度为96.97%其中包括8706.5 M/Z和8579.2M/Z(t值分别为2.562和2.783,P值均<0.05),经分离鉴定其为阿朴脂蛋白AⅡ. 结论 阿朴脂蛋白AⅡ是肝癌的差异蛋白之一,可能与肝癌的发生有关.

The screening and identification of Apolipoprotein A-II from serum differential proteins in hepatocellular carcinoma patients

Objective To screen differential proteins in serum from hepatocellular carcinoma (HCC) patients by Proteomic Technology and to purify and identify them. Methods Surface enhanced laser desorp-tion Ionization time of flight-mass spectrum (SELDI-TOF-MS) was employed to screen differential proteins in serum from 33 HCC patients and 33 control cases, and then to purify and identify them using isoelectric precipitation, Tricine sodium dodecyl sulphate ployacrylamide gel electrophoresis (Tricine-SDS-PAGE) and high performance liquid chromatography tandem Mass Spectrum (HPLC-MS). Result 65 protein peaks in the range of relative molecular weight from 2000 to 10 000 were found significant difference (P < 0.05) between the patient group and control group. Based on these differential protein peaks, diagnostic model for HCC detection was established and its sensitivity and specificity were 100% and 96.97 % respectively. Proteins with 8706.5 and 8579.2 relative molecular weights (the t value was 2.562 and 2.783 respectively, and P value was 0.013 and 0.015 respectively) out of the 65 differential proteins were purified and identified, and then recognized as Apolipoprotein A Ⅱ (Apo AⅡ). Conclusion Apo A II is probably a differential protein of HCC and maybe related to the pathogenesis of HCC.