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该研究对不同转移能力的人肝癌细胞系全细胞蛋白进行差异蛋白质组分析,以期发现与肝癌转移相关的候选分子。选择具有高、低转移能力的人肝癌细胞系HCCLM6和MHCC97H为研究材料,提取细胞全蛋白,FASP酶切、iTRAQ标记后进行质谱鉴定和定量分析,并利用Uni Prot数据库和GOfact软件对差异蛋白进行生物信息学分析,包括亚细胞定位、生物过程和分子功能富集分析。共鉴定了5 033种蛋白质,其中5 013种蛋白质有定量信息,发现91种差异蛋白(|ratio|≥1.5,单样本t检验,P<0.05),其中39种蛋白在高转移细胞HCCLM6中上调,52种下调。差异蛋白的细胞定位主要为细胞质和细胞膜,GO分析显著富集到细胞黏附生物过程和蛋白结合生物功能。该研究建立了高低转移能力的人肝癌细胞系HCCLM6和MHCC97H细胞全蛋白的差异表达谱,发现了4种新的与肝癌转移相关的候选蛋白(微管蛋白链β-2B、着丝粒蛋白F、层黏连蛋白亚基α5和囊泡相关膜蛋白5),为进一步研究肝癌转移机制提供了重要的数据参考。
In this study, differential proteomic analysis of whole cell proteins of human hepatocellular carcinoma cell lines with different metastatic potential was carried out in order to find candidate molecules related to metastasis of liver cancer. The human hepatoma cell lines HCCLM6 and MHCC97H with high and low metastatic potential were selected as the research material, and the whole cell protein, FASP digestion and iTRAQ were extracted and identified by mass spectrometry. The differential proteins were analyzed by Uni Prot database and GOfact software Bioinformatics analysis, including subcellular localization, biological processes and molecular function enrichment analysis. A total of 5 033 proteins were identified, among which 5 013 proteins had quantitative information and 91 proteins (| ratio | ≥1.5, one-sample t-test, P <0.05) were found, of which 39 proteins were upregulated in highly metastatic HCCLM6 , 52 kinds of down. The cellular localization of the differentially expressed proteins was mainly cytoplasm and cell membrane, and GO analysis was significantly enriched in cell adhesion bio-process and protein binding biological function. This study established differential expression profiles of total proteins in human hepatocellular carcinoma cell lines HCCLM6 and MHCC97H with high and low metastatic potential and found four new candidate proteins related to metastasis of liver cancer (tubulin chain β-2B, centromere protein F , Laminin subunit α5 and vesicle-associated membrane protein 5) provide important data for further study on the mechanism of liver cancer metastasis.