Quantitative proteome analysis of HCC cell lines with different metastatic potentials by SILAC
Quantitative proteome analysis of HCC cell lines with different metastatic potentials by SILAC
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SILAC对不同转移潜能HCC细胞系的定量蛋白质组分析
DOI：
10.1002/pmic.200800280
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发表时间：
2008-12-01
期刊：
影响因子：
3.4
通讯作者：
He, Fuchu
中科院分区：
文献类型：
--
作者：
Chen, Ning;Sun, Wei;He, Fuchu
肝细胞癌（HCC）是全球最常见的癌症之一，转移是HCC治疗失败和高死亡率的主要原因。为了进一步探讨HCC转移的机制，寻找候选的诊断标志物和治疗靶点，采用细胞培养氨基酸稳定同位素标记（SILAC）技术对具有低和高转移潜能的HCC细胞系MHCC97L和HCCLM6进行差异蛋白质组分析。使用LTQ-FT质谱总共鉴定了2335个可靠的蛋白质，其中91个蛋白质在HCCLM6中上调，61个蛋白质下调。大多数上调的蛋白参与粘附、形态发生和脂质合成，而许多下调的蛋白参与电子传输，这可能对 HCC 转移至关重要。通过细胞系中的蛋白质印迹验证了六种失调蛋白。有趣的是，通过Western blotting进一步证实了培养物上清液中溶质载体家族12成员2（SLC 12A2）和蛋白质二硫键异构酶A4（PDIA4）的上调，并通过ELISA进一步证实了具有不同转移潜能的HCC患者血清中溶质载体家族12成员2（SLC 12A2）和蛋白质二硫键异构酶A4（PDIA4）的上调。我们的研究不仅为 HCC 转移机制提供了有价值的见解，而且还为预测 HCC 转移提供了潜在的候选生物标志物。
Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and metastasis is the main cause for treatment failure and high fatality of HCC. In order to make further exploration into the mechanism of HCC metastasis and to search for the candidates of diagnostic marker and therapeutic target, stable‐isotope labeling by amino acids in cell culture (SILAC) technique was employed to conduct differential proteome analysis on HCC cell lines – MHCC97L and HCCLM6 with low and high metastatic potentials. In total, 2335 reliable proteins were identified using LTQ‐FT mass spectrum, among which 91 proteins were upregulated and 61 proteins were downregulated in HCCLM6. Most of the upregulated proteins were involved in adherence, morphogenesis, and lipid synthesis, while lots of the downregulated proteins were involved in electron transport, which might be crucial for HCC metastasis. Six dysregulated proteins were validated by Western blotting in the cell lines. Interestingly, the upregulation of solute carrier family 12 member 2 (SLC 12A2) and protein disulfide‐isomerase A4 (PDIA4) were further confirmed in the culture supernatants by Western blotting and in the sera of HCC patients with different metastatic potentials by ELISA. Our study provided not only the valuable insights into the HCC metastasis mechanisms but also the potential candidate biomarkers for prediction of HCC metastasis.