Article ID Journal Published Year Pages File Type
1225922 Journal of Proteomics 2014 12 Pages PDF
Abstract

•iTRAQ-labeling and SWATH™ quantification proteomic were performed to analyze the NSCLC cell secretome.•iTRAQ-labeling proteomics analyze the early-stage and late-stage pooled NSCLC serum.•High expression of CD109 in NSCLC tissue and serum suggested that it was associated with NSCLC progression and metastasis.

To identify cancer-related proteins, we used isobaric tags in a relative and absolute quantitation (iTRAQ) proteomic approach and SWATH™ quantification approach to analyze the secretome of an isogenic pair of highly metastatic and low metastatic non-small-cell lung cancer (NSCLC) cell lines. In addition, we compared two groups of pooled serum samples (12 early-stage and 12 late-stage patients) to mine data for candidates screened by iTRAQ-labeled proteomic analysis. A total of 110 proteins and 71 proteins were observed to be significantly differentially expressed in the cell line secretome and NSCLC sera, respectively. Among these proteins, CD109 was found to be highly expressed in both the highly metastatic cell line secretome and the group of late-stage patients. A sandwich ELISA assay also demonstrated an elevation of serum CD109 levels in individual NSCLC patients (n = 30) compared with healthy subjects (n = 19). Furthermore, CD109 displayed higher expression in lung cancer tissues compared with their matched noncancerous lung tissues (n = 72). In addition, the knockdown of CD109 influenced several NSCLC cell bio-functions, for instance, depressing cell growth, affecting cell cycle phases. These phenomena suggest that CD109 plays a critical role in NSCLC progression.Biological significanceWe simultaneously applied two quantitative proteomic approaches—iTRAQ-labeling and SWATH™—to analyze the secretome of metastatic cell lines, in order to explore the cancer-associated proteins in conditioned media. In this study, our results indicate that CD109 plays a critical role in non-small-cell lung cancer (NSCLC) progression, and is overexpressed in advanced NSCLC.

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Related Topics
Physical Sciences and Engineering Chemistry Analytical Chemistry
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