体外培养的不同亚型肝癌细胞差异蛋白的初步筛查

THE PRELIMINARY ANALYSIS OF DISTINCT PROTEINS FOR DIFFERENT TYPES OF LIVER CANCER GELL LINES

目的 分析体外培养的不同亚型肝癌细胞株蛋白质表达差异 ,筛选肝癌的标志蛋白。 方法 采用表面增强激光解吸离子化 (SELDI)蛋白质芯片技术检测了两种肝癌细胞株 (肝细胞癌 ,SMMC 772 1和肝内胆管细胞癌 ,IHCCC 981 0 )以及正常肝细胞 (L1 0 2 )的蛋白质谱。选用阴离子交换柱处理上述细胞总蛋白 ,用 6种不同pH的洗脱液洗脱柱床 ,通过改变洗脱液的pH值将总蛋白分成不同pI范围的 6组。每个组分用WCX2和IMAC3两种芯片检测。采用PBSII C型蛋白质芯片阅读机读取数据 ,获得的数据采用Ciphergen公司的ProteinchipSoftware3.0 .2软件分析。 结果 与L0 2细胞相比 ,有 1 2个蛋白质在两种肝癌细胞株中均出现明显的变化 ,其中 4个差异蛋白在肝癌细胞中高表达 ,8个差异蛋白在肝癌细胞中低表达。在对两种肝癌细胞株的蛋白质谱比较时发现 ,不同亚型的肝癌细胞有其各自的标志蛋白 ,其中 1 1个蛋白分子在SMMC 772 1细胞中高表达 ;在IHCCC 981 0细胞中 7个蛋白分子高表达 ,1 0个蛋白分子低表达。 结论 不同亚型肝癌细胞与正常肝细胞蛋白质谱比较时有共同的差异蛋白 ,同时不同亚型肝癌细胞之间的蛋白质谱表达也有差别。

Objective To discover the common biomarkers in liver cancer cell lines and detect the distinct proteins for different types of liver cancer cell lines. Methods We performed surface enhanced laser desorption ionization(SELDI)mass spectrometry using protein chip technology to compare the differentially expressed proteins in two types of liver cancer cell lines(hepatocellular carcinoma cell line,SMMC7721 and intrahepatic cholangiocellular carcinoma cell line,IHCCC9810)with normal liver cell line(L02).Anionexchange columns were used to fractionate the total protein of these cells with 6 designated pH washing solutions.Two different types of Protein Chip arrays,IMACCu and WCX2,were employed.Protein chips were examined in PBSIIC Protein Chip Reader(Ciphergen Biosystem Inc)and the protein profiling was analyzed by Proteinchip Software 3.0.2. Results The results showed that 12 distinct proteins were found in two types of liver cancer cell lines with 4 proteins upregulated and 8 proteins downregulated when compared with L02 cell line.Furthermore,the changes of expression in different types of liver cancer cells were found with 11 biomarkers upregulated in hepatocellular carcinoma cells,and 7 biomarkers upregualted and 10 biomarkers downregulated in intrahepatic cholangiocellular carcinoma cell.Conclusion The results suggest that there are same distinct proteins in protein profiling between liver cancer cells and normal liver cells,and the discriminated proteins between different types of liver cancer cells are also found.