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. 2017 Feb 11;8(3):479-489.
doi: 10.7150/jca.17192. eCollection 2017.

Identifying DCN and HSPD1 as Potential Biomarkers in Colon Cancer Using 2D-LC-MS/MS Combined with iTRAQ Technology

Guoqing Li  1 , Maoyu Li  2 , Xujun Liang  2 , Zhefeng Xiao  2 , Pengfei Zhang  2 , Meiying Shao  2 , Fang Peng  2 , Yongheng Chen  2 , Yuanyuan Li  3 , Zhuchu Chen  2
Affiliations

Identifying DCN and HSPD1 as Potential Biomarkers in Colon Cancer Using 2D-LC-MS/MS Combined with iTRAQ Technology

Guoqing Li et al. J Cancer. .

Abstract

Colon cancer is one of the most common types of gastrointestinal cancers and the fourth cause of cancer death worldwide. To discover novel diagnostic biomarkers for colon cancer and investigate potential mechanisms of oncogenesis, quantitative proteomic approach using iTRAQ-tagging and 2D-LC-MS/MS was performed to characterize proteins alterations in colon cancer and non-neoplastic colonic mucosa (NNCM) using laser capture microdissection-harvested from the two types of tissues, respectively. As a result, 188 DEPs were identified, and the differential expression of two DEPs (DCN and HSPD1) was further verified by Western blotting and immunohistochemistry. KEGG pathway analysis disclosed that the DEPs were related to signaling pathways associated with cancer; furthermore, DCN and HSPD1 are in the relative central hub position among protein-protein interaction subnetwork of the DEPs. The results not only shed light on the mechanism by the DEPs contributed to colonic carcinogenesis, but also showed that DCN and HSPD1 are novel potential biomarkers for the diagnosis of colon cancer.

Keywords: DCN; HSPD1; Proteomics analysis; carcinogenesis.; colon cancer; iTRAQ.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

Figures

Figure 1
Figure 1
Classification of differentially expressed proteins by gene ontology biological process (A), molecular function (B), and subcellular component (C).
Figure 2
Figure 2
Mass spectrometry (MS) spectra of precursor ions in consistent with peptides are part of DCN and HSPD1. (A) (left) The sequence ETADTDTADQVMASFK allows the identification of DCN; (right) the released iTRAQ reporter ions provide the relative quantitation of DCN from the two tissues evaluated. (B) (left) The sequence ALMLQGVDLLADAVAVTMGPK allows the identification of HSPD1; (right) the released iTRAQ reporter ions provide the relative quantitation of HSPD1 from the two tissues evaluated. Non-neoplastic colonic mucosa, labeled with iTRAQ reagent 115,117; colon cancer, labeled with iTRAQ reagents 114,116.
Figure 3
Figure 3
Expressional levels of DCN and HSPD1 in non-neoplastic colonic mucosa (NNCM) (N) and colon cancer (C). (left) Representative results of Western blotting analysis of DCN and HSPD1 in NNCM (N) and colon cancer (C). (right) Histogram shows the expression levels of the two proteins in these tissues as determined by densitometric analysis. β-actin was used as a loading control. Columns, mean from 10 cases of tissues; bars, S.D.
Figure 4
Figure 4
Representative results of immunohistochemistry of DCN and HSPD1 in tissue specimens. (A) DCN-non-neoplastic colonic mucosa. (B) DCN-colon cancer. (C) HSPD1-non-neoplastic colonic mucosa (D) HSPD1-colon cancer. Main image, ×100.
Figure 5
Figure 5
Significant KEGG pathways that are involved in protein-protein interaction network of differentially expressed proteins.
Figure 6
Figure 6
Protein-protein interaction networks of differentially expressed proteins. Opaque red circles, HSPD1 and DCN of nodes.
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