title: Proteomic analysis on N, N'-dinitrosopiperazine-mediated metastasis of nasopharyngeal carcinoma 6-10B cells.
abstract: Nasopharyngeal carcinoma (NPC) has a high metastatic feature. N,N'-Dinitrosopiperazine (DNP) is involved in NPC metastasis, but its mechanism is not clear. The aim of this study is to reveal the pathogenesis of DNP-involved metastasis. 6-10B cells with low metastasis are from NPC cell line SUNE-1, were used to investigate the mechanism of DNP-mediated NPC metastasis.
  
  
  
  6-10B cells were grown in DMEM containing 2H4-L-lysine and 13C 6 15 N4-L-arginine or conventional L-lysine and L-arginine, and identified the incorporation of amino acid by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Labeled 6-10B cells were treated with DNP at 0 -18 μM to establish the non-cytotoxic concentration (NCC) range. NCC was 0 -10 μM. Following treatment with DNP at this range, the motility and invasion of cells were detected in vitro, and DNP-mediated metastasis was confirmed in the nude mice. DNP increased 6-10B cell metastasis in vitro and vivo. DNP-induced protein expression was investigated using a quantitative proteomic. The SILAC-based approach quantified 2698 proteins, 371 of which showed significant change after DNP treatment (172 up-regulated and 199 down-regulated proteins). DNP induced the change in abundance of mitochondrial proteins, mediated the status of oxidative stress and the imbalance of redox state, increased cytoskeletal protein, cathepsin, anterior gradient-2, and clusterin expression. DNP also increased the expression of secretory AKR1B10, cathepsin B and clusterin 6-10B cells. Gene Ontology and Ingenuity Pathway analysis showed that DNP may regulate protein synthesis, cellular movement, lipid metabolism, molecular transport, cellular growth and proliferation signaling pathways.
  
  
  
  DNP may regulate cytoskeletal protein, cathepsin, anterior gradient-2, and clusterin expression, increase NPC cells motility and invasion, is involved NPC metastasis.
authors.affiliation: Zhuhai Hospital, Jinan University, Zhuhai, Guangdong, China.
authors.forename: Yuejin
authors.initials: Y
authors.lastname: Li
authors.forename: Na
authors.initials: N
authors.lastname: Liu
authors.forename: Damao
authors.initials: D
authors.lastname: Huang
authors.forename: Zhenlin
authors.initials: Z
authors.lastname: Zhang
authors.forename: Zhengke
authors.initials: Z
authors.lastname: Peng
authors.forename: Chaojun
authors.initials: C
authors.lastname: Duan
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Tang
authors.forename: Gongjun
authors.initials: G
authors.lastname: Tan
authors.forename: Guangrong
authors.initials: G
authors.lastname: Yan
authors.forename: Wenhua
authors.initials: W
authors.lastname: Mei
authors.forename: Faqing
authors.initials: F
authors.lastname: Tang
mesh_terms: D000818:Animals; D002277:Carcinoma; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003598:Cytoskeletal Proteins / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling; D053858:Metabolic Networks and Pathways; D051379:Mice; D008819:Mice, Nude; D000077274:Nasopharyngeal Carcinoma; D009303:Nasopharyngeal Neoplasms / Q000378:metabolism / Q000473:pathology; D009362:Neoplasm Metastasis; D009602:Nitrosamines / Q000494:pharmacology*; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects; D020543:Proteome / Q000187:drug effects*; D053719:Tandem Mass Spectrometry
chemical_list: D003598:Cytoskeletal Proteins; D009602:Nitrosamines; D020543:Proteome; C026156:N,N'-dinitrosopiperazine