Proteomic differential display analysis identified upregulated astrocytic phosphoprotein PEA‐15 in human malignant pleural mesothelioma cell lines
Proteomic differential display analysis identified upregulated astrocytic phosphoprotein PEA‐15 in human malignant pleural mesothelioma cell lines
复制标题
蛋白质组差异显示分析发现人恶性胸膜间皮瘤细胞系中星形细胞磷蛋白 PEA-15 上调
作者：
Y. Kuramitsu;Hiromu Miyamoto;Toshiyuki Tanaka;Xiulian Zhang;M. Fujimoto;K. Ueda;Toshiki Tanaka;K. Hamano;K. Nakamura
应用双向凝胶电泳法和LC-MS/MS联用技术对人恶性胸膜间皮瘤细胞系和人胸膜间皮细胞系进行了蛋白质组差异显示分析。人恶性胸膜间皮瘤细胞系为NCI-H28、NCI-H2052和NCI-H2452，人胸膜间皮细胞系为MET-5A。在MET-5A和NCI-H2052之间，从2-DE的结果中我们发现了38个表达水平不同的蛋白质点，其中NCI-H2052的28个蛋白质点高于MET-5A，其他10个蛋白质点低于MET-5A。用LC-MS/MS分析了这些斑点，并用肽序列标签进行了鉴定。然而，从其他细胞系的2-DE结果来看，在所有三种MPM细胞系中只有一种持续上调的蛋白，即星形细胞磷酸蛋白PEA-15。用针对PEA-15的特异性抗体进行的Western blotting证实，与MET-5A细胞和患者的正常胸膜组织相比，PEA-15在所有三种MPM细胞系中的表达水平都升高。PEA-15在NCI-H2052细胞中表达下调，与阴性对照细胞相比，PEA-15沉默的NCI-H2052细胞的增殖抑制7-15%。这些结果提示PEA-15的表达可能与MPM的肿瘤发生有关。
We performed proteomic differential display analysis of human malignant pleural mesothelioma (MPM) cell lines and a human pleural mesothelial cell line by using 2‐DE and LC‐MS/MS. The human MPM cell lines were NCI‐H28, NCI‐H2052 and NCI‐H2452, and the human pleural mesothelial cell line was MeT‐5A. Between MeT‐5A and NCI‐H2052, we found 38 protein spots whose expression levels were different, from the results of 2‐DE; 28 protein spots appeared higher, and 10 other protein spots lower in NCI‐H2052 than in MeT‐5A. These spots were analyzed by LC‐MS/MS analysis and identified by a peptide sequence tag. However, from the results of 2‐DE of the other cell lines, there was only one consistently upregulated protein, astrocytic phosphoprotein PEA‐15, in all three MPM cell lines. Western blotting using specific antibodies against PEA‐15 confirmed the elevated expression level of PEA‐15 in all three MPM cell lines compared with MeT‐5A cells and normal pleura tissues from patients. PEA‐15 was knocked down in NCI‐H2052 cells, and the proliferation of PEA‐15‐silenced NCI‐H2052 cells was suppressed 7–15% compared with negative control cells. These results suggest that PEA‐15 expression is likely to be associated with the tumorigenesis of MPM.
影响因子：
11.8
作者：
Formstecher, E;Ramos, JW;Chneiweiss, H
通讯作者：
Chneiweiss, H
影响因子：
11.2
作者：
Jagadeeswaran, R;Ma, PC;Salgia, R
通讯作者：
Salgia, R