Abstract
To identify metastasis-related proteins in nasopharyngeal carcinoma (NPC), iTRAQ-tagging combined with 2D LC-MS/MS analysis was performed to identify the differentially expressed proteins (DEPs) in high metastatic NPC 5-8F cells and non-metastatic NPC 6-10B cells, and qRT-PCR and Western blotting were used to confirm DEPs. As a result, 101 DEPs were identified by proteomics, and 12 DEPs were selectively validated. We further detected expression of three DEPs (RAN, SQSTM1 and TRIM29) in a cohort of NPC tissue specimens to assess their value as NPC metastatic biomarkers, and found that combination of RAN, SQSTM1 and TRIM29 could discriminate metastatic NPC from non-metastatic NPC with a sensitivity of 88% and a specificity of 91%. TRIM29 and RAN expression level were closely correlated with lymph node and distant metastasis and clinical stage (P <0.05) in NPC patients. Finally, a combination of loss-of-function and gain-of-function approaches was performed to determine the effects of TRIM29 on NPC cell proliferation, migration, invasion and metastasis. The results showed that TRIM29 knockdown significantly attenuated while TRIM29 overexpression promoted NPC cell in vitro proliferation, migration and invasion and in vivo metastasis. The present data first time show that SQSTM1, RAN and TRIM29 are novel potential biomarkers for predicting NPC metastasis, demonstrate that TRIM29 is a metastasis-promoted protein of NPC.
Highlights
Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in southern China and Southeast Asia
A total of 862 non-redundant proteins were repeatedly identified by Isobaric tags for relative and absolute quantitation (iTRAQ) labeling and 2D-LCMS/MS analyses in nasopharyngeal carcinoma (NPC) cells, 79.37% of which were identified with ≥2 peptides matches
ITRAQ labeling combined with 2D-LC-MS/MS was used to identify differentially expressed proteins (DEPs) in the NPC cells with different metastatic potentials
Summary
Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in southern China and Southeast Asia. Metastasis is one of distinctive characteristics of NPC [1]. NPC is sensitive to radiotherapy, significant rates of relapse and distant metastasis still occur after therapy, which is a major cause for NPC lethality [2]. The five-year survival rate following combined treatment with radiotherapy and adjuvant cisplatin chemotherapy is 50-60% and the rates of fiveyear cumulative local relapse and distant metastasis are 2030 and 20-25%, respectively [3]. The molecular mechanism of NPC metastasis is not yet well-defined. To develop better diagnosis and treatment approaches, it is important to identify proteins related to NPC metastasis and understand the molecular basis of NPC metastasis
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