Abstract
In our previous study, five different secretory proteins, including GSN, ADAMTSL4, CALR, PPIA and TXN, have been identified to be associated with the nasopharyngeal carcinoma (NPC) metastasis. In this work, the 5 proteins were further investigated. Bioinformatics analysis suggested that they might play an important role in the process of NPC development. Western blotting analysis showed that all of these 5 targets could be secreted into extracellular by both high metastatic NPC 5-8F cells and non-metastatic NPC 6-10B cells. Except for GSN, the expressions of ADAMTSL4, CALR, PPIA and TXN proteins in extracts of the 5-8F and 6-10B cells were significantly different (P < 0.05). Thus, the expressions of these 4 differentially expressed proteins were further tested in a cohort of NPC tissue specimens. The results indicated that the expression levels of ADAMTSL4 and TXN were highly correlated with the lymph node and distant metastasis (P<0.05) in NPC patients. Moreover, Enzyme-linked immunosorbent assay (ELISA) was used to investigate the concentrations of the ADAMTSL4 and TXN in serum specimens of NPC patients. The results revealed that serum ADAMTSL4 expression level was closely correlated with lymph node metastasis and clinical stage (P<0.05) in NPC patients, and it was able to discriminate metastasis NPC from non-metastasis NPC with a sensitivity of 75.6% and a specificity of 64.7%. The present data show for the first time that the ADAMTSL4 and TXN may be novel and potential biomarkers for predicting the NPC metastasis.Furthermore, the serum ADAMTSL4 could be a potential serum tumor biomarker for prognosis of NPC.
Highlights
Nasopharyngeal carcinoma (NPC) is a common head and neck cancer in southern China and Southeast Asia [1,2,3]
Supplementary Figrue S1B showed the miRNAs of genes corresponding to the 5 differentially expressed proteins (DEPs).the GenMAPP pathway analysis showed that the 5 DEPs significantly enriched in unfolded protein binding signaling pathway, cell proliferation pathway, cell motility pathway, etc.They may play an important role in the process of nasopharyngeal carcinoma development as Supplementary Figrue S1C described
To check the qualities of the cell extracts (CE) and conditioned media (CM), Western blotting analysis was conducted to examine the presence of two abundant cytoplasmic proteins: β-actin and glyceraldehyde-3phosphate dehydrogenase (GAPDH)
Summary
Nasopharyngeal carcinoma (NPC) is a common head and neck cancer in southern China and Southeast Asia [1,2,3]. To develop better diagnosis and treatment approaches, it is important to identify the proteins related to the NPC metastasis, especially the proteins from human serum or other body fluids. The secretome, defined by Tjalsma and his colleagues [8] in 2000, is used to describe proteins secreted/shed from cells, tissues or organs. It plays an important role in growth, angiogenesis, invasion www.impactjournals.com/oncotarget and metastasis of cancer-cells. Through screening and identifying the secretome in tumor cells, many researchers are trying to discover tumor biomarkers and therapeutic targets from human serum or other body fluids using proteomics technologies, and have received great achievements [9,10,11]
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