In this study, we aim to identify
biomarkers for
gastric cancer metastasis using a quantitative proteomics approach. The
proteins extracted from a panel of 4
gastric cancer cell lines, two derived from primary
cancer (AGS, FU97) and two from
lymph node metastasis (AZ521, MKN7), were labeled with iTRAQ (8-plex)
reagents and analyzed by 2D-LC-MALDI-TOF/TOF MS. In total, 641
proteins were identified with at least a 95% confidence. Using cutoff values of >1.5 and <0.67, 19
proteins were found to be up-regulated and 34 were down-regulated in the metastatic versus primary
gastric cancer cell lines respectively. Several of these dysregulated
proteins, including
caldesmon, were verified using Western blotting. It was found that
caldesmon expression was decreased in the two
metastasis-derived cell lines, and this was confirmed by further analysis of 7
gastric cancer cell lines. Furthermore, immunohistochemical staining of 9 pairs of primary
gastric cancer and the matched
lymph node metastasis tissue also corroborated this observation. Finally, knockdown of
caldesmon using
siRNA in AGS and FU97
gastric cancer cells resulted in an increase in cell migration and invasion, while the overexpression of
caldesmon in AZ521 cells led to a decrease in cell migration and invasion. This study has thus established the potential role of
caldesmon in
gastric cancer metastasis, and further functional studies are underway to delineate the underlying mechanism of action of this
protein.