As an in vitro model for type II human lung cancer, A549 cells resist cytotoxicity via phosphorylation of proteins as demonstrated by many studies. However, to date, no large-scale phosphoproteome investigation has been conducted on A549. Here, we performed a systematical analysis of the phosphoproteome of A549 by using mass spectrometry (MS)-based strategies. This investigation led to the identification of 337 phosphorylation sites on 181 phosphoproteins. Among them, 67 phosphoproteins and 230 phosphorylation sites identified appeared to be novel with no previous characterization in lung cancer. Based on their known functions as reported in the literature, these phosphoproteins were functionally organized into highly interconnected networks. Western blotting and immunohistochemistry analyses were performed to validate the expression of a bottleneck phosphoprotein YAP1 in cancer cell lines and tissues. This dataset provides a valuable resource for further studies on phosphorylation and lung carcinogenesis.

这篇文章，打了A549细胞的磷酸化谱，总体上分析了一下。最后通过网络分析，找到candidate，作了验证。

构建网络之后，进行了一些拓扑分析：

最后通过betweenness的分析，鉴定网络中的bottleneck分子：

选取其中一个bottleneck分子YAP1进行了Westernblot和免疫组化验证，结果均具有统计学显著性。

细胞样本的westernblot用ANOVA分析，p = 0.01046；组织样本的westernblot用Welch two sample t test， p = 0.01969；IHC用Wilcoxon signed rank test，p = 0.03301。