肝癌发生不同病理阶段血清蛋白质组的比较

目的 比较肝癌发生不同病理阶段血清蛋白质组的表达变化情况,从中寻找特异性标志物.方法 用双向凝胶电泳分离正常人,慢性肝炎、肝硬化和肝癌患者的血清蛋白质,结合质谱技术对差异点进行鉴定; Western blot检测结合珠蛋白β链以验证蛋白质水平的表达.计量资料数据以均数±标准差(x±s)表示,采用方差分析和LSD检验进行两两比较.结果 4组血清的双向电泳图谱经软件匹配分析,并与基质辅助激光解析电离飞行时间质谱相结合,成功鉴定出结合珠蛋白,SAA1、SP40等30个差异蛋白质点,K cluster聚类分析不同的变化模式.在差异蛋白质点中,7个蛋白质点均为结合珠蛋白,呈现由正常→肝炎→肝硬化持续下调、到肝癌又上调的模式.Western blot证实结合珠蛋白β链的表达与双向电泳表达相一致.结论 在肝癌发生的动态过程中各疾病阶段的血清蛋白质差异表达有4种明显变化模式,可能为肝癌的早期诊断和预后提供依据,与肝癌发生发展相关的结合珠蛋白很可能是肝硬化发展到肝癌的一个潜在早期诊断标志物.

Comparison of the serum proteomes of pathological stages during hepatocarcinogenesis

Objectives To compare the 2-DE profiles for serum proteins of different pathological stages during hepatocareinogenesis. Methods Sera from hepatoeellular carcinoma patients, cirrhosis patients,chronic hepatitis patients and healthy controls were collected. After sonication, albumin and immunogiobulin (IgG) depletion, and desalination, sera were subjected to 2-DE, the differential protein spots were identified by MALDI-TOF-MS. Western blot was used to validate these differentially expressed proteins. Results 2-DE sera protein profiles were obtained fi'om the patient suffering from HCC, liver cirrhosis, chronic hepatitis,healthy controls in each group. From optimized 2-DE gel images of the above groups, 96 protein spots with more than 2-fold difference in intensity between the two groups were selected by image master 6.0 software,differential proteins including haptoglobin, SAAI and SP40 were identified by MALDI-TOF-MS/MS. 7 different spots within more than 30 protein spots belonged to the same haptogiobin family. The differential expression of haptoglobin was confirmed by western blot. Conclusions Four protein expression patterns have been identified during the pathological stages of hepatocarcinogenesis. Haptoglobin is significantly increased from liver cirrhosis to HCC. It implies that haptoglobin might be a potential biomarker in the early diagnosis of liver cancer.