Proteomic analysis on N, N[prime]-dinitrosopiperazine-mediated metastasis of nasopharyngeal carcinoma 6-10B cells


Nasopharyngeal carcinoma (NPC) has a high metastatic feature. N,N[prime]-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.

Results: 6-10B cells were grown in DMEM containing 2H4-L-lysine and 13C615N4-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 muM to establish the non-cytotoxic concentration (NCC)range. NCC was 0 -10 muM.

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.

Conclusion: DNP may regulate cytoskeletal protein, cathepsin, anterior gradient-2, and clusterin expression, increase NPC cells motility and invasion, is involved NPC metastasis.

Author: Yuejin LiNa LiuDamao HuangZhenlin ZhangZhengke PengChaojun DuanXiaowei TangGongjun TanGuangrong YanWenhua MeiFaqing Tang
Credits/Source: BMC Biochemistry 2012, 13:25



Published on: 2012-11-19



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