乙腈预处理血清寻找肝细胞癌发病相关的蛋白质

【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｡

Proteomic Screening of Serum Hepatocellular Carcinoma-associated Proteins by an Acetonitrile-pretreatment Method

Objective]To apply an acetonitrile-pretreatment method for proteomic analysis of serum of patient with hepatocellular carcinoma(HCC),searching for the proteins that might be related to occurrence and development of HCC.Methods]Serum samples were recruited from 12 patients with primary HCC and 12 health adult.Firsdy,the sera were pretreated with acetonitrile (ACN),and the high-abundance proteins were removed,then two-dimensional electrophoreaia(2-DE)was performed to screen the differentially expressed proteins between the patient with HCC and health control.Finally,the differentially expressed proteins in the 2-DE gel were identified by a MALDI-TOF/TOF mass spectrometry.Results]After the sera were pretreated with 0%,20% and 30% ACN,2-DE analysis showed that the number of the protein spots in the gels were 532±96,623±102,and 674±123,respectively.By the proteomic analysis of the ACN-pretreatment sera of patient with HCC and health control,we found that tetraiodo-L-thyronine and proapolipoproteinin were up-regulated,vitamin D-binding protein precurser and transferrin were downregulated in the HCC serum.Conclusion]Proteomic analysis of serum using the ACN-pretreatment method can improve the detection of proteins which were non-specifically binding to the albumin.Abnormal expression of tetraiodo-L-thyronine,proapolipoproteinin,vitamin D-binding protein precursor and transferrin in the serum are related to HCC.