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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 analyze the differences of protein expression in different subtypes of hepatocellular carcinoma cell lines cultured in vitro and screen the marker proteins of hepatocellular carcinoma. Methods Two types of hepatocellular carcinoma cell lines (hepatocellular carcinoma, SMMC772 1 and intrahepatic cholangiocarcinoma, IHCCC 981 0) and normal hepatocytes (L1 0 2) were examined by surface enhanced laser desorption ionization (SELDI) Protein spectrum. The total cellular protein was treated with an anion exchange column. The column was eluted with 6 different pH eluents. The total protein was divided into 6 groups with different pI by changing the pH of the eluate. Each component was tested with both WCX2 and IMAC3 chips. Using PBSII C-type protein chip reader to read the data, the data obtained using Ciphergen’s Proteinchip Software3.0 .2 software analysis. Results Compared with L0 2 cells, 12 proteins showed obvious changes in both hepatocellular carcinoma cell lines. Four differential proteins were highly expressed in hepatocellular carcinoma cells and eight differential proteins were low in hepatoma cells. When comparing the protein profiles of the two hepatoma cell lines, it was found that the liver cancer cells of different subtypes had their own marker proteins, of which 11 protein molecules were highly expressed in SMMC 772 1 cells and 7 proteins in IHCCC 981 0 cells Molecular high expression, 10 protein molecules are low expression. Conclusion The protein profiles of hepatoma cells in different subtypes are different from those in normal liver cells. There are also differences in the protein profiles between different subtypes of HCC cells.