肝病良恶性腹腔积液前期处理后差异蛋白质组学分析

Analysis of malignant and nonmalignant ascites after sample preparations in liver disease by differential expression of proteins

目的 分析肝炎后肝硬化及其相关性原发性肝癌患者腹腔积液前期处理后差异蛋白质组学,探讨肝病癌性腹腔积液具有特征性的标志蛋白组合模式.方法 应用表面加强激光解析电离飞行时间质谱（SELDI-TOF-MS）技术,对46例肝炎后肝硬化（肝硬化腹腔积液组）及27例肝炎后肝硬化相关性原发性肝癌患者（肝癌腹腔积液）腹腔积液前期处理后蛋白质进行检测,对蛋白质波谱进行分析,并寻找合理运算规则以确定能够区分原发性肝癌和肝炎后肝硬化腹腔积液患者的蛋白质组学图谱.结果 浓缩前肝硬化腹腔积液组的蛋白质浓度低于肝癌腹腔积液组[（8 ±4）g/L比（39±4） g/L],差异有统计学意义（P＜0.05）.浓缩后,肝硬化腹腔积液组蛋白质浓度与肝癌腹腔积液组的蛋白浓度接近[（54±5）g/L比（58±5）g/L],差异无统计学意义（P＞0.05）.离心前,肝癌腹腔积液组腹腔积液细胞计数明显多于肝硬化腹腔积液组[（3146±1362）×10^6/L比（287 ±51）×10^6/L] （P＜0.05）;离心1次后,肝硬化腹腔积液组腹腔积液细胞计数为（9 ±3）×10^6/L,肝癌腹腔积液组腹腔积液细胞计数为（19 ±7）×10^6/L,2组差异无统计学意义（P＞0.05）;离心2次后,2组腹腔积液中细胞计数均为0.在质荷比为11 467、4 475时,肝癌腹腔积液组特征峰表达值均高于肝硬化腹腔积液组,7 852时,肝癌腹腔积液组特征峰表达值低于肝硬化腹腔积液组.SELDI-TOF-MS技术检测肝癌与肝硬化的灵敏度为81.5％（22/27）,特异性为87.0％（40/46）.结论 腹腔积液前期处理能满足SELDI-TOF-MS技术的要求,确立的标志蛋白组合模式的特异性及敏感度较高.

Objective To screen ascites biomarkers in posthepatitis cirrhosis and its relation to primary hepatic carcinoma patients with surface enhanced laser desorption /ionization time of flight mass spectrometry （SELDI-TOF-MS） technique.Methods Proteinic spectra was generated by SELDI-TOF-MS technique.The ascites from 46 posthepatitis cirrhosis（cirrhosis ascites group） and its related to 27 primary hepatic carcinoma patients （hepatic carcinoma ascites group）were analyzed for selection of the specific protein of liver cancer.The biomarker patterns were used.Results Before contration,the level of protein in cirrhosis ascites group was less than that in hepatic carcinoma ascites group [（8 ± 4） g/L vs （39 ± 4） g/L,P 〈 0.05] ; after concentration,the level of protein in cirrhosis ascites group was close to the level in hepatic carcinoma ascites group[（54 ±5）g/L vs （58 ±5）g/L,P 〉 0.05].Before centrifugation,the cell count of ascites in carcinoma ascites group was more than that in cirrhosis ascites group[（3 146 ± 1 362） × 10^6/L vs （287 ± 51） × 10^6/L] （P 〈 0.05） ; the cell count of ascites in carcinoma ascites group and cirrhosis ascites group were 0.At the m/z was 4 475 and 11 467,the characterissic protein expression peak level was higher in carcinoma ascites group than that in cirrhosis ascites group（P 〈0.05）,while at the m/z was 7 852,the characteristic protein expression peak level was less than that of cirrhosis ascites group （P〈0.05）.The diagnosis way was established with the sensitivity of 81.5% （22/27） and the specificity of 87.0% （40/46）.Conclusions Sample preparations for ascites proteome can meet the demands of surface enhanced laser desorption/ionization time of flight mass spectrometry.The characteristic protein expression peak has higher sensitivity and specificity.Differential expression protein method has important significance for further study of the ascites proteomic changes in the development and progression of liver cancer.