肝细胞肝癌门静脉癌栓相关蛋白表达研究

Proteomic analysis on portal vein tumor thrombus-associated proteins for hepatocellular carcinoma

目的应用比较蛋白组学研究与肝细胞肝癌门静脉癌栓形成相关的蛋白质分子标记物。方法利用双向凝胶电泳(2-DE)对同一患者的肝癌原发灶组织和其门静脉癌栓组织进行总蛋白的分离,两组之间的差异蛋白通过质谱分析和蛋白数据库搜索鉴定,并用免疫印迹(Western 印迹)法对差异蛋白点进行进一步检测验证。结果共鉴定出20有意义的蛋白质。12个蛋白仅在肝癌原发灶组织表达或表达明显增高,其中包括半乳糖苷凝-1(Galectin-1)、高迁移率族蛋白-1(HMGBI)、过氧化氧化还原蛋白1(peroxiredoxin 1)、亲环蛋白 B(Cyclophilin B)、增殖细胞核抗原(PCNA)等,同时我们发现8个蛋白仅门静脉癌栓中表达或者高表达包括膜联蛋白5(Annexin V)、TriosephosphateIsomerase。对于门静脉癌栓组织高表达的 Annexin V,应用 Western 印迹在蛋白水平上验证了2-DE 结果。结论肝癌门静脉癌栓形成与多种蛋白质相关,其中 Annexin V 高表达可能发挥重要作用。

Objective A comparative proteomic approach was used to analyze proteins relevant to portal vein tumor thrombus forming. Methods proteins extracted from five pairs of matched primary tumor/ tumor thrombus samples in the same patient were separated by two-dimensional gel electrophoresis （2-DE）. Selected proteins exhibiting statistically significant alternations were identified by mass spectrometry. Western blotting was further performed to examine the expression of the candidate proteins. Result There were 20 significant proteins were identified in total, Among the 20 spots, 12 proteins were up-regulated proteins in primary tumor tissue, including Galectin-1, HMGBI, peroxiredoxin l, Cyclophilin B, PCNA. whereas 8 were up-regulated proteins in tumor thrombus samples, including Annexin V, Triosephosphate Isomerase. Western blotting Confirmed the difference of Annexin V on protein level. Conclusion There are many proteins associated with the formation of PVTT in HCC. The overexpression of Annexin V may serve as a biomarker for early detection and therapeutic targets to HCC with PVTT.