Identification and Functional Validation of Caldesmon as a Potential Gastric Cancer Metastasis-associated Protein

卡尔德蒙 癌症 转移 癌症研究 癌细胞 污渍 蛋白质组学 细胞培养 生物 细胞迁移 病理 细胞 分子生物学 化学 医学 生物化学 基因 遗传学 钙调蛋白
作者
Qingzhi Hou,Hwee Hoon Tan,Kiat Hon Lim,Teck Kwang Lim,Avery Khoo,Iain Beehuat Tan,Khay Guan Yeoh,Maxey C. M. Chung
出处
期刊:Journal of Proteome Research [American Chemical Society]
卷期号:12 (2): 980-990 被引量:51
标识
DOI:10.1021/pr3010259
摘要

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.
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