二亚硝基哌嗪上调AGR2促进鼻咽癌细胞转移

doi:10.3969/j.issn.1004-616x.2016.02.007

Abstract

Abstract: OBJECTIVE: To investigate theinduction of AGR2 expression by dinitrosopiperazine (DNP) in nasopharyngeal cancer cell (NPC) 6-10B and to explore involvement of DNP in NPC metastasis. METHODS: Non-cytotoxic concentrations of DNP on 6-10B cells were determined using MTT. Expression of AGR2 protein and mRNA in 6-10B cells which were treated with DNP were detected by indirect immunofluorescence, Western blotting and real-time RT-PCR, respectively. In addition, silencing the expression of AGR2 by siRNA-AGR2 was investigated. The capabilities of invasion and migration of 6-10B cells were investigated using Transwell assay. RESULTS: The range of non-cytotoxic concentrations (NCC) of DNP was 0-10.0 μmol/L. The results of indirect immunofluorescence showed AGR2 mainly expressed in cytoplasm after DNP treatment. These and 5-8F cells had stronger fluorescence intensity than untreated 6-10B cells. The DNP-induced AGR2 expression showed dose- and time-dependent effects. Real-time RT PCR analyses showed that AGR2 mRNA expression rate was 1.01±0.08 in the untreated 6-10B cells and 3.96±0.15 in the cells treated with 8.0 μmol/L DNP. The difference was significant. In addition, the relative fold gene expression of AGR2 in 5-8F cells was higher than that in non-treated 6-10B cells (P<0.05). The transwell migration and invasion data showed that siRNA-AGR2 transfection blocked AGR2 expression in 6-10B cells. In addition, both DNP-induced invasion and motility were dramatically decreased when AGR2 expression was blocked (P<0.05). However, DNP could effectively induce cell invasion (P<0.05) and motility (P<0.05) when AGR2 was not blocked. CONCLUSION: DNP could up-regulate AGR2 expression in 6-10B cells through transcriptional regulation mechanism and then mediate the metastasis of nasopharyngeal cancer cells.

Key words: nasopharyngeal cancer, dinitrosopiperazine, AGR2, neoplasm metastasis

CLC Number:

R73-37

WANG Weiwei, TANG Faqing, LI Yuejin. Dinitrosopiperazine-mediated up-regulation of AGR2 expression promotes metastasis of nasopharyngeal cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(2): 115-120.

References

[1] Wei WI,Sham JST. Nasopharyngeal carcinoma[J]. Lancet, 2005,365(9476):2041-2054.
[2] Feng BJ,Huang W,Shugart YY,et al. Genome-wide scan for familial nasopharyngeal carcinoma reveals evidence of linkage to chromosome 4[J]. Nat Genet,2002,31(4):395-399.
[3] Yu MC,Henderson BE. Intake of Cantonese-style salted fish as a cause of nasopharyngeal carcinoma[J]. IARC Sci Publ, 1987,(84):547-549.
[4] Yu MC,Nichols PW,Zou XN,et al. Induction of malignant nasal cavity tumours in Wistar rats fed Chinese salted fish[J]. Br J Cancer,1989,60(2):198-201.
[5] 湖南医学院肿瘤研究所. 亚硝胺类化合物诱发大鼠鼻咽癌的实验研究[J]. 科学通报,1978,23(12):756-769.
[6] 杨大俊,区宝祥. 二亚硝基哌嗪(DNP)体外诱导正常人胚鼻咽成纤维细胞转化的研究[J]. 癌症,1988,7(6):417-420.
[7] Tang F,Jiang H,Duan Z,et al. Profile of telomerase and telomerase RNA expression in nasopharyngeal carcinogenesis of rats induced by N,N'dinitrosopiperazine (DNP)[J]. Chin J Pathol,2001,30(2):125-128.
[8] Aberger F,Weidinger G,Grunz H,et al. Anterior specification of embryonic ectoderm:the role of the Xenopus cement gland-specific gene XAG-2[J]. Mech Dev,1998, 72(1/2):115-130.
[9] Kumar A,Godwin JW,Gates PB,et al. Molecular basis for the nerve dependence of limb regeneration in an adult vertebrate[J]. Science:New York,2007,318(5851):772-777.
[10] Loong HH,Ma BB,Chan AT. Update on the management and therapeutic monitoring of advanced nasopharyngeal cancer[J]. Hematol Oncol Clin North Am,2008,22(6):1267-1278.
[11] Sham JS,Cheung YK,Chan FL,et al. Nasopharyngeal carcinoma:pattern of skeletal metastases[J]. Br J Radiol, 1990,63(747):202-205.
[12] Tang FQ,Duan CJ,Huang DM,et al. HSP70 and mucin 5B:novel protein targets of N,N'-dinitrosopiperazine-induced nasopharyngeal tumorigenesis[J]. Cancer Sci,2009,100(2):216-224.
[13] Tang F,Zou F,Peng Z,et al. N,N'-Dinitrosopiperazine-mediated Ezrin protein phosphorylation via activation of Rho kinase and protein kinase C is involved in metastasis of nasopharyngeal carcinoma 6-10B cells[J]. J Biol Chem,2011, 286(42):36956-36967.
[14] Li Y,Liu N,Huang D,et al. Proteomic analysis on N,N'-dinitrosopiperazine-mediated metastasis of nasopharyngeal carcinoma 6-10B cells[J]. BMC Biochem,2012,13(1):25.
[15] Vivekanandan P,Micchelli ST,Torbenson M. Anterior gradient-2 is overexpressed by fibrolamellar carcinomas[J]. Hum Pathol,2009,40(3):293-299.
[16] Ramachandran V,Arumugam T,Wang H,et al. Anterior gradient 2 is expressed and secreted during the development of pancreatic cancer and promotes cancer cell survival[J]. Cancer Res,2008,68(19):7811-7818.
[17] Pizzi M,Fassan M,Balistreri M,et al. Anterior gradient 2 overexpression in lung adenocarcinoma[J]. Appl Immunohistochem Mol Morphol,2012,20(1):31-36.
[18] Innes HE,Liu D,Barraclough R,et al. Significance of the metastasis-inducing protein AGR2 for outcome in hormonally treated breast cancer patients[J]. Br J Cancer,2006,94(7):1057-1065.
[19] Fritzsche FR,Dahl E,Pahl S,et al. Prognostic relevance of AGR2 expression in breast cancer[J]. Clin Cancer Res,2006, 12(6):1728-1734.

Dinitrosopiperazine-mediated up-regulation of AGR2 expression promotes metastasis of nasopharyngeal cancer cells

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[2]	WANG Lu1;3;REN Xing2; AO Lin3; PANG Xue-li2; ZHANG Li-shi1; CAO Jia3;. DNA Damage in Nasopharyngeal Cancer Patients Undergoing Radiotherapy [J]. , 2009, 21(4): 268-270.
[3]	YU Dong-hong;GUO Bing-qing;CHENG Ze-nong;et al. Expression of MMP-2 and its Correlation with Tumor Proliferative Activity and Tumor Metastasis in Human Esophodeal Carcinoma [J]. , 2004, 16(2): 83-85.