论文部分内容阅读
【目的】应用乙腈(ACN)预处理血清的方法,对肝细胞癌(HCC)患者的血清进行蛋白质组分析,寻找与HCC发病相关的蛋白质。【方法】收集原发性HCC患者和健康人的血清各12例,首先使用乙腈预处理,然后去除白蛋白和免疫球蛋白等高丰度蛋白质,再进行双向电泳(2-DE)分析,筛选HCC与健康对照血清中有显著性差异的蛋白质斑点,并进行MALDI-TOF/TOF质谱鉴定。【结果】血清经0%、20%和30%浓度的乙腈预处理后,进行2-DE分析发现蛋白质斑点的检出数量分别为532±96、623±102和674±123;对预处理后HCC患者和正常人的血清进行差异分析,发现甲状腺素(Tetraiodo-L-Thyronine)和Proapolipoprotein的表达上调,而维生素D结合蛋白(Vitamin D-binding Protein)和铁传递蛋白(Transferrin)的表达下调。【结论】对血清样本的蛋白质组分析,应用乙腈预处理会增加与白蛋白非特异性结合的蛋白质的检出;HCC患者血清中甲状腺素等4个蛋白的表达异常与HCC相关。
【Objective】 Serum samples from patients with hepatocellular carcinoma (HCC) were pretreated with acetonitrile (ACN) for proteomic analysis to find proteins related to the pathogenesis of HCC. 【Methods】 Serum samples of 12 patients with primary HCC and healthy individuals were collected. Twelve samples were pre-treated with acetonitrile and then excised high-abundance proteins such as albumin and immunoglobulin, followed by two-dimensional electrophoresis (2-DE) HCC and healthy control serum were significantly different protein spots, and MALDI-TOF / TOF mass spectrometry identification. 【Results】 The result of 2-DE analysis showed that the number of detected protein spots were 532 ± 96, 623 ± 102 and 674 ± 123 respectively after the serum was pretreated with 0%, 20% and 30% acetonitrile. After pretreatment HCC patients and normal serum were analyzed for differences and found that the expression of Tetraiodo-L-Thyronine and Proapolipoprotein were up-regulated, while the expression of Vitamin D-binding protein and Transferrin were down-regulated. 【Conclusion】 Proteome analysis of serum samples using acetonitrile pretreatment will increase the detection of proteins that do not specifically bind to albumin. The abnormal expression of four proteins such as thyroxine in HCC patients is associated with HCC.