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Proteome analysis of hepatocellular carcinoma cell strains, MHCC97-H and MHCC97-L, with different metastasis potentials

机译:具有不同转移潜能的肝癌细胞株MHCC97-H和MHCC97-L的蛋白质组分析

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摘要

To better understand the mechanism underlying hepatocellular carcinoma (HCC) metastasis and to search for potential markers for HCC prognosis, differential proteome analysis on two HCC cell strains with high and low metastatic potentials, MHCC97-H and MHCC97-L, was conducted using two-dimensional (2-D) gel electrophoresis followed by matrix-assisted laser desorption/time of flight mass spectrometry and liquid chromatography ion trap mass spectrometry. Image analysis of silver-stained 2-D gels revealed that 56 protein spots showed significant differential expression in MHCC97-H and MHCC97-L cells (Student's t-test, P < 0.05) and 4 protein spots were only detected in MHCC97-H cells. Fourteen protein spots were further identified using in-gel tryptic digestion, peptide mass fingerprinting and tandem mass spectrometry. The expressions of pyruvate kinase M2, ubiquitin carboxy-terminal hydrolase L1, laminin receptor 67 kDa, S100 calcium-binding protein A4, thioredoxin and cytokeratin 19 were elevated in MHCC97-H cells. However, manganese superoxide dismutase, calreticulin precursor, cathepsin D, lactate dehydrogenase B, non-metastatic cell protein 1, cofilin 1 and calumenin precursor were down-regulated in MHCC97-H cells. Intriguingly, most of these identified proteins have been reported to be associated with tumor metastasis. The functional implications of alterations in the levels of these proteins are discussed.
机译:为了更好地了解肝细胞癌(HCC)转移的潜在机制并寻找可能的肝癌预后标志物,使用两种方法对两种具有高和低转移潜能的HCC细胞株MHCC97-H和MHCC97-L进行差异蛋白质组分析维(2-D)凝胶电泳,然后进行基质辅助激光解吸/飞行时间质谱和液相色谱离子阱质谱。银染二维凝胶的图像分析显示,在MHCC97-H和MHCC97-L细胞中有56个蛋白斑点显示出显着差异表达(Student's t检验,P <0.05),仅在MHCC97-H细胞中发现了4个蛋白斑点。 。使用凝胶内胰蛋白酶消化,肽质量指纹图谱和串联质谱进一步鉴定了14个蛋白斑点。在MHCC97-H细胞中,丙酮酸激酶M2,泛素羧基末端水解酶L1,层粘连蛋白受体67 kDa,S100钙结合蛋白A4,硫氧还蛋白和细胞角蛋白19的表达升高。然而,在MHCC97-H细胞中,锰超氧化物歧化酶,钙网蛋白前体,组织蛋白酶D,乳酸脱氢酶B,非转移性细胞蛋白1,cofilin 1和钙粘蛋白前体被下调。有趣的是,据报道,大多数这些鉴定出的蛋白质与肿瘤转移有关。讨论了这些蛋白质水平改变的功能含义。

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