title: Identification of nasopharyngeal carcinoma metastasis-related biomarkers by iTRAQ combined with 2D-LC-MS/MS.
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.
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Long
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Facai
authors.initials: F
authors.lastname: Cui
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Lepan
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Tao
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wu
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Manlin
authors.initials: M
authors.lastname: Xiang
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Yunlai
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Shiyang
authors.initials: S
authors.lastname: Qiu
authors.affiliation: The Higher Educational Key Laboratory for Cancer Proteomics and Translational Medicine of Hunan Province, Xiangya Hospital, Central South University, Changsha, Hunan, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Xiao
authors.affiliation: Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Yi
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D002277:Carcinoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002851:Chromatography, High Pressure Liquid; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D057166:High-Throughput Screening Assays; D006801:Humans; D008207:Lymphatic Metastasis; D008297:Male; D008875:Middle Aged; D000077274:Nasopharyngeal Carcinoma; D009303:Nasopharyngeal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D009361:Neoplasm Invasiveness; D009367:Neoplasm Staging; D011237:Predictive Value of Tests; D040901:Proteomics / Q000379:methods*; D034622:RNA Interference; D015203:Reproducibility of Results; D000071456:Sequestosome-1 Protein / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D020931:ran GTP-Binding Protein / Q000378:metabolism*
chemical_list: D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; C120587:RAN protein, human; C099718:SQSTM1 protein, human; D000071456:Sequestosome-1 Protein; C087296:TRIM29 protein, human; D014157:Transcription Factors; D020931:ran GTP-Binding Protein
keywords: RAN; SQSTM1; TRIM29; iTRAQ; metastasis; nasopharyngeal carcinoma