肝癌门静脉癌栓相关小分子的比较蛋白质组学分析

目的 筛选肝细胞肝癌门静脉癌栓(PVTT)形成过程中起重要作用的低相对分子质量蛋白质标记物.方法 选取肝癌门静脉癌栓和肝癌无癌栓患者原发瘤组织各12例,利用16%SDS-PAGE和Tris-Tricine系统进行双向电泳,得到小相对分子质量蛋白表达图谱,经Image Master软件对比分析后,基质辅助激光解析飞行时间质谱(MALDI-TOF MS/MS)对差异点进行鉴定.免疫印迹法对鉴定蛋白进行初步验证.结果 无PVTT和PVTT的原发瘤组织间共检测到88个差异点,其中相对分子质量(MW)低于20×103的差异点有42个.与无PVTT组比较,PVTT组中有41个蛋白点表达上调(其中19个MW低于20×103),47个蛋白点表达下调(23个MW低于20×103).经质谱鉴定共发现钙结合蛋白S100A11,脂肪酸结合蛋白-1等11种胶内差异小分子蛋白质.免疫印迹验证结果证实S100A11在有或无PVTT的原发瘤组织中的确存在表达差异.结论 PVTT的原发瘤组织与无PVTT原发瘤组织中存在差异表达的低相对分子质量蛋白,推测肝癌侵袭转移性与多种差异蛋白相关,S100A11可能是其中一种.

Comparative proteome analysis of PVTT related small molecules

Objective To screen low molecular weight protein biomarkers which played an im-portant role during the course of portal vein tumor thrombi (PVTT) formation in hepatocellular carcinoma (HCC). Methods We first selected 2 groups ( each group had 12 cases ) of primary tumor tissue from human HCC with or without PVTT. Then a stable and standard low MW ( < 20×103 ) protein electropho-resis platform was sued by changing both SDS-PAGE gel concentration and components of electrophoretic buffer system in the second step of 2-DE. Using optimized 2-DE and MALDI-TOF MS/MS, differentially expressed proteomes were compared and studied in the 2 groups. By using Western blot, the results were i-dentified. Results Protein samples from PVTT group or non-PVTT group ran 2-DE for 3 times repeatedly under the same conditions. Eighty-eight differential spots were detected between primary tumor tissues of the two groups. Compared with non-PVTT group, there were 41 up-regulated spots (including 19 MW lower than 20×103 ) and 47 down-regulated spots in PVTT group ( including 23 MW lower than 20×103). Thir-ty-six spots were picked and redundancy was eliminated. Finally, 11 significantly differential low MW pro-teins such as S100A11 and fat acid binding (FAB) protein etc were further identified by MALDI-TOF MS/MS. The results from Western blotting validation confirmed the differential expression of S100A11 in primary tumor tissue with PVTT or without PVTT. Conclusion The low MW protein profile of HCC pri-mary tumor tissue with PVTT displayed obvious difference compared with HCC tissue without PVTT. The results indicated that the invasive and metastatic characteristic of HCC bad relations with many differential-ly expressed proteins,including S100A11.