Open AccessTissue‐based quantitative proteomics to screen and identify the potential biomarkers for early recurrence/metastasis of esophageal squamous cell carcinoma
Author(s) -
Cai XuWei,
Yu WeiWei,
Yu Wen,
Zhang Qin,
Feng Wen,
Liu MiNa,
Sun MengHong,
Xiang JiaQing,
Zhang YaWei,
Fu XiaoLong
Publication year - 2018
Publication title -
cancer medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.403
H-Index - 53
ISSN - 2045-7634
DOI - 10.1002/cam4.1463
Subject(s) - metastasis , proteomics , biomarker , blot , cancer research , immunohistochemistry , cell growth , tissue microarray , cell , cell culture , gene knockdown , biology , esophageal cancer , cancer , pathology , microbiology and biotechnology , medicine , gene , biochemistry , genetics
Esophageal squamous cell carcinoma ( ESCC ) is the eighth cause of cancer‐related deaths worldwide. To screen potential biomarkers associated with early recurrence/metastasis (R/M) of ESCC patients after radical resection, ESCC patients were analyzed by a comparative proteomics analysis using iTRAQ with RPLC ‐ MS to screen differential proteins among R/M groups and adjacent normal tissues. The proteins were identified by qRT ‐ PCR , Western blotting, and tissue microarray. The protein and mRNA expression difference of PHB 2 between tumor tissues of ESCC patients and adjacent normal tissues, ESCC patients with and without metastasis, four ESCC cell lines and normal esophageal epithelial cells were inspected using immunohistochemical staining, qRT ‐ PCR , and Western blotting. The EC 109 and TE 1 cells were used to establish PHB 2 knockdown cell models, and their cell proliferation and invasion ability were determined by cell counting method, Transwell ® assay. Thirteen proteins were selected by cutoff value of 0.67 fold for underexpression and 1.5‐fold for overexpression. Seven proteins were confirmed to be associated with R/M among the 13 proteins. The potential biomarker PHB 2 for early recurrence/metastasis of ESCC was identified. PHB 2 expression was related to the OS of ESCC patients ( P = 0.032) and had high levels in the tumor tissues and human cell lines of ESCC ( P = 0.0002). Also, the high PHB 2 expression promoted the metastasis of ESCC ( P = 0.0075), suggesting high PHB 2 expression was a potential prognostic biomarker. Experiments showed that PHB 2 could significantly promote the proliferation and cell invasion ability of human ESCC cell lines and the knockdown of PHB 2 suppressed the phosphorylation level of AKT , as well as the expression of MMP 9 and RAC 1. PHB 2 could predict the early metastasis of ESCC patients.