HepG2细胞蛋白质组双向电泳条件的优化与图谱的建立

Establishment and Optimization of Two-dimensional Gel Electrophoresis Technique for Proteomics of HepG2 Cells

目的：优化双向电泳的样品处理方法,建立HepG2细胞蛋白质双向电泳图谱。方法：用含10%胎牛血清的RPM11640培养液培养HepG2细胞,胰酶消化后收集细胞,加入细胞裂解液使细胞充分裂解,冰浴超声离心后取上清液进行双向凝胶电泳,电泳图谱经考马斯亮蓝染色及图像扫描与分析。结果：通过对HepG2细胞蛋白提取液进行双向电泳,在13 cm pH 3~10（L）IPG胶条上可分离到（297±23）个重复性及分辨率均较好的蛋白质点,蛋白斑点的匹配率为83%。结论：HepG2细胞蛋白质组双向电泳条件的优化及图谱的建立,为进一步研究肝癌蛋白质组学奠定了基础。

Objective：To optimize the treatment methods of Two-dimensional Electrophoresis sample and establish the 2-DE profiles of proteome of HepG2 cells.Methods： HepG2 cells were cultured in RPMI 1640 culture medium containing 10% fetal bovine serum,then harvested after digestion by trypsin,and lysed in lysis buffer.After centrifugation,supernatant was separated by two-dimensional gel electrophoresis,stained by Coomassie Brilliant Blue,and then followed by image scanning and analysis.Results：（297±23）protein spots were detected in the 2-DE profiles on 13 cm IPG strip（pH 3～10 L） of HepG2 cells proteome separated by 2-D PAGE.A 83% match rate of protein spots between gels was obtained.Conclusion：The establishment and optimization of two-dimensional gel electrophoresis technique for proteomics of HepG2 cells laid foundation for further research on hepatocellular carcinoma proteomics.