肺腺癌SPC-A1细胞双向电泳图谱的优化

Optimization of 2-D electrophoresis map of pulmonary adenocarcinoma cell strain SPC-A1

双向电泳技术+质谱技术已经成为蛋白组学研究的基本策略之一。获得细胞或组织经过某种处理前后的双向电泳图谱,然后经专业2-D图像分析软件找到差异表达(有无的差异或是表达量的差异)的蛋白质斑点,再经过质谱分析鉴定这些蛋白质的种类,可以全面、系统地研究该处理因素对总蛋白质表达的影响。而要想准确地获得某种组织或细胞的双向电泳图谱,建立与之兼容的方法必不可少。本文通过使用和SPC—A1细胞株兼容的总蛋白质抽提方法,采用不同pH梯度的IPG(immorbilized pH gradient)干胶条,获得了较为清晰的蛋白质斑点,并重点分离了特定局部区域的蛋白质,优化了本细胞株的双向电泳图谱,为进一步分析药物处理前后的总蛋白质表达差异创造了条件。

Two-dimensional electrophoresis combined with mass spectrotometry is one of the basic strategy of proteomic research. Different 2-D electrophoresis map of certain cell or tisssue are acquired before and after some kind of treatment.Subsequently, professional 2-D analysis software is used to locate the different protein spots, followed by analysis of mass spectrotometry to i-dentify and characterize them. Thus, we could study the influence of this treatment on the expression of total protein expression comprehensively and systematically. Building a compatible 2-D elctrophoresis method is necessary to accquire an accurate 2-D map of certain cell or tissue. In this article, we accquired a clear and high-resolution 2-D map by using total protein extraction method compatible with SPC-A1 cell train. We also separated and visualized proteins of certain region intensively by applying IPG strips of different pH range. By all these efforts made, we have optimized the 2-D map of this cell train and paved the way for further analysis.