人肝细胞性肝癌组织转移相关分子的比较蛋白质组学研究

目的 应用比较蛋白质组学方法研究肝细胞性肝癌(HCC)转移相关的关键蛋白分子。方法 用双向凝胶电泳(2DE)对有转移的HCC组织和未发生转移的HCC组织总蛋白进行分离，两组间差异蛋白点用质谱和数据库搜索鉴定，并在蛋白质和mRNA水平上进一步检测验证。结果 鉴定出l6个差异表达蛋白，包括S100钙结合蛋白(S100)、热休克蛋白27(HSP27)、细胞角蛋白18(CK18)等。对在转移性HCC组织中表达显著增高的HSP27，应用western blot分析在蛋白水平上验证了2DE结果，RT-PCR检测出两组问mRNA水平也存在差异，但不显著，免疫组织化学检测显示HSP27主要定位于肝细胞胞浆内。结论 HCC转移与多种蛋白表达相关，其中HSP27高表达可能在转移中发挥作用，可能为潜在的判断HCC转移的分子标记或控制转移的治疗靶点。

Proteomic analysis on metastasis-associated proteins of hepatocellular carcinoma tissues

Objective A comparative proteomic approach was used to identify and analyze proteins relevant to metastasis of hepatocellular carcinoma (HCC). Methods Proteins extracted from 12 liver tumor tissue specimens (6 with metastases and 6 without) were separated by two-dimensional gel electrophoresis (2-DE). Comparative analyses of 2-DE protein patterns between the two groups were done using computerized image analysis. Selected proteins exhibiting statistically significant alternations were identified by mass spectrometry. Immunohistochemistry, Western blotting and RT-PCR were performed to examine the expressions of the candidate proteins. Results 16 proteins including HSP27, S100A11, CK18 were identified using mass spectrometry, which were related to cell mobility, signal transduction, and energy metabolism respectively. Of these, HSP27 was found to be uniquely over-expressed in 2-DE maps of all metastatic HCCs when compared to the non-metastatic HCC tissues. Immunohistochemistry and Western blotting of HCC tissues confirmed this difference while RT-PCR did not. Conclusion There are different proteins working together that affect the metastasis of HCCs. The overexpression of HSP27 may serve as a biomarker for early detection and therapeutic targets to the metastatic phenotype of HCC. The role of HSP27 in HCC metastasis warrants further investigation.