蛋白质芯片飞行质谱技术检测体外培养的肝癌细胞株与转染HBV的肝癌细胞株蛋白质的差异表达

Identification of protein differently expressed between hepatoma cell line and Hepatitis B Virus-transfected hepatoma cell line

目的:运用SELDI蛋白质芯片技术检测体外培养的肝癌细胞株(HepG2)与转染乙肝病毒的肝癌细胞株(HepG2.2.15)蛋白质的差异表达,从而为进一步研究肝癌的发病机制奠定基础. 方法:常规培养上述两种细胞,细胞状态良好时收集细胞,裂解细胞后采用SELDI—TOF—MS技术用WCX2 芯片检测HepG2、HepG2.2.15蛋白质组学的差异. 结果:WCX2芯片共捕获91个蛋白,其中HepG2、HepG 2.2.15细胞差异蛋白共19个,9个蛋白在HepG2 细胞中表达量增高,10个蛋白在HepG2细胞中表达量降低.通过在Swiss蛋白数据库中搜索,发现Mr 11 081 蛋白与钙结合蛋白S100 A10相符,其他几种蛋白暂时寻找不到. 结论:SELDI蛋白芯片技术检测肝癌细胞株与转染乙肝病毒的肝癌细胞株蛋白质的差异表达方法简便,敏感性高,重复性好,本文发现的这些组织特异性蛋白生物标记对我们从血清或组织标本中筛选和鉴定不同型别肝癌细胞之间的标志蛋白有重要意义.

AIM： To detect the different protein expression between Hepatitis B Virus （HBV）-transfected hepatoma cell line （HepG2.2.15） and its parental cell line （HepG2） in vitro using the surfase-enhenced laser desorption/ionization （SELDI） protein chip.METHODS： HepG2 and HepG2.2.15 cell lines were cultured by routine method and then collected. All the cells were lysed when they were in good conditions, and the protein expression in the lysate was detected by WCX2 chip using surfase-enhenced laser desorption/ionization-time-of-flight mass spectrometry （SELDI-TOF-MS）. RESULTS： Ninety-one proteins were captured by WCX2 chip, of which nineteen were differently expressed between HepG2.2.15 and HepG2 cells. Among the nineteen proteins, nine were up-regulated and ten were down-regulated in HepG2 cells. After search in SWISS-PROT, the protein with Mr11081 was found accordant to calcium-binding protein S100A10, and the other ones were not confirmed yet. CONCLUSION： SELDI protein chip platform is simple,sensitive and repeatable in the detection of differently expressed proteins, whose specific biological markers play important roles in screening and identifying the marker proteins of the cells from different types of liver cancer, between hepatoma cell line and HBV-transfected hepatoma cell line.