Proteomic differential display analysis identified upregulated astrocytic phosphoprotein PEA-15 in human malignant pleural mesothelioma cell lines.
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.
Kuramitsu
Yasuhiro
Y
Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi, Japan. climates@yamaguchi-u.ac.jp
Miyamoto
Hiromu
H
Tanaka
Toshiyuki
T
Zhang
Xiulian
X
Fujimoto
Masanori
M
Ueda
Kazuhiro
K
Tanaka
Toshiki
T
Hamano
Kimikazu
K
Nakamura
Kazuyuki
K
D051017:Apoptosis Regulatory Proteins; D001253:Astrocytes / Q000378:metabolism*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic / Q000528:radiation effects; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008654:Mesothelioma / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010997:Pleural Neoplasms / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D015854:Up-Regulation
D051017:Apoptosis Regulatory Proteins; D047908:Intracellular Signaling Peptides and Proteins; C092039:PEA15 protein, human; D010750:Phosphoproteins