Differential protein expression in EC304 gastric cancer cells induced by alphastatin.
OBJECTIVE
To explore the differential protein expression profile in EC304 gastric cancer cells induced by alphastatin.


METHODS
Cultured EC304 cells in the exponential phase of growth were randomly divided into alphastatin and control groups. Total proteins were extracted and the two dimensional electrophoresis (2-DE) technique was applied to analyze differences in expression with ImageMaster 2D Platinum 5.0 software. Proteins were identified using the MASCOT database and selected differently expressed proteins were characterised by western blotting and immunofluorescence.


RESULTS
1350 ± 90 protein spots were detected by the ImageMaster software in the 2-DE gel images from the control and alphastatin groups. The match rate was about 72-80% for the spectrum profiles, with 29 significantly different protein spots being identified, 10 upregulated, 16 downregulated, two new and one lost. The MASCOT search scores were 64-666 and the peptide matching numbers were 3-27 with sequence coverage of 8-62%. Twenty-three proteins were checked by mass spectrometry, including decrease in Nm23 and profilin-2 isoform b associated with the regulation of actin multimerisation induced by extracellular signals.


CONCLUSIONS
The proteome in EC304 cells is dramatically altered by alphastatin, which appears to play an important role in modulating cellular activity and anti-angiogenesis by regulating protein expression and signal transduction pathways through Nm23 and profilin-2 isoform b, providing new research directions for anti-angiogenic therapy of gastric cancer.
Department of General Surgery, Chinese People's Liberation Army General Hospital, Beijing, China.
Xin-Xin
XX
Wang
Rong-Ju
RJ
Sun
Meng
M
Wu
Tao
T
Li
Yong
Y
Zhang
Lin
L
Chen
D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D002478:Cells, Cultured; D015536:Down-Regulation / Q000187:drug effects; D005340:Fibrinogen / Q000008:administration & dosage / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D054778:NM23 Nucleoside Diphosphate Kinases / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects; D051304:Profilins / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D013274:Stomach Neoplasms / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects; D062948:rho Guanine Nucleotide Dissociation Inhibitor alpha / Q000187:drug effects / Q000378:metabolism
D020533:Angiogenesis Inhibitors; D054778:NM23 Nucleoside Diphosphate Kinases; C495741:PFN2 protein, human; D051304:Profilins; C514422:alphastatin; D062948:rho Guanine Nucleotide Dissociation Inhibitor alpha; D005340:Fibrinogen; C517938:NME1 protein, human