摘要
目的:利用蛋白质组学方法,识别并鉴定幽门螺杆菌(Helicobacterpylori,H.pylori)处理HepG2细胞后差异表达的蛋白质,为进一步探讨幽门螺杆菌对肝细胞的病理作用机制奠定基础。方法:运用固相pH梯度双向凝胶电泳技术分离H.pylori处理前后HepG2细胞的总蛋白质,用图象分析软件比较分析,以识别差异表达的蛋白质;应用质谱仪(massspectrometry)得到相应的肽质指纹图谱(peptidemassfingerprint,PMF),然后搜索数据库鉴定差异蛋白质。结果:鉴定出12种差异表达蛋白质,这些蛋白质包括整合素β1、蛋白激酶Cα、PINCH蛋白、Ras相关蛋白Rab37、LIM同源盒蛋白Lhx1、HLAⅠ类抗原、血管抑制素和金属蛋白酶抑制因子2等。这些差异蛋白质涉及基因表达调控、细胞免疫、细胞生长和信号传导等众多关键事件。结论:H.pylori处理前后HepG2的蛋白质组具有差异,这些差异表达的蛋白质可能参与了H.pylori对肝细胞的病理作用过程,这种蛋白质组的差异分析有助于进一步研究H.pylori与人类肝脏疾病的关系。
OBJECTIVE： To investigate the pathological effect of Helicobacter. pylori on human hepatic cells, proteomic methods were used to find and identify differential expression proteins in HepG2 cells that were induced by H. pylori treatment. METHODS： The total proteins of untreated and H. pylori treated HepG2 cells were separated by immobilized pH gradient （ IPG ）-based two-dimensional gel electrophoresis （ 2-DE ）. The differential expression proteins were analyzed by using image analysis software, then identified by using mass spectrometry and database searching. RESULTS： Twelve differential proteins were successfully identified. These proteins included integrin β-1, protein kinase C alpha, PINCH protein, Ras-related protein Rab-37, LIM/homeobox protein Lhx1, HLA class Ⅰ histocompatibility antigen, angiostatin, metalloproteinase inhibitor 2 etc. , which involved in transcription regulation, metabolism, signaling pathway and so on. CONCLUSIONS： There is a significant difference at protein level between untreated and H. pylori treated HepG2. These proteins may be involved in the pathological process of H. pylori on HepG2. It suggests that the differential expression analysis of proteomes may be useful to further study on the relation of H. pylori and human liver diseases.
出处
《肿瘤防治杂志》
2005年第21期1609-1614,共6页
China Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(30271171)
国家教育部优秀骨干教师基金(教技司2000-65)
湖南省自然科学基金(01jjy2114)
湖南省卫生厅科研基金(00022)
