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

癌变·畸变·突变 ›› 2016, Vol. 28 ›› Issue (2): 115-120.doi: 10.3969/j.issn.1004-616x.2016.02.007

二亚硝基哌嗪上调AGR2促进鼻咽癌细胞转移

王巍巍, 唐发清, 李跃进

珠海市人民医院/暨南大学附属珠海医院检验科, 广东珠海 519000

收稿日期:2015-11-20 修回日期:2016-01-20 出版日期:2016-03-31 发布日期:2016-03-31
通讯作者: 李跃进,E-mail:liyuejincat@163.com E-mail:liyuejincat@163.com
作者简介:王巍巍,E-mail:wangweiweicat@163.com
基金资助:
国家自然科学基金资助项目(81402265)

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

WANG Weiwei, TANG Faqing, LI Yuejin

Department of Clinical Laboratory, Zhuhai People's Hospital & Zhuhai Hospital of Jinan University, Zhuhai 519000, Guangdong, China

Received:2015-11-20 Revised:2016-01-20 Online:2016-03-31 Published:2016-03-31

摘要/Abstract

摘要： 目的: 探讨二亚硝基哌嗪(DNP)通过调控AGR2表达参与鼻咽癌转移的分子机制。方法: 以具有低转移潜能的鼻咽癌细胞6-10B作为研究材科,应用四甲基噻唑蓝(MTT)法检测DNP对6-10B细胞的非毒性浓度;分别采用间接免疫荧光法、Western blot和实时荧光定量PCR法检测DNP对6-10B细胞中AGR2蛋白及mRNA表达的影响;采用针对AGR2基因的特异性小干扰RNA(siRNA)沉默6-10B细胞AGR2的表达,并用Transwell小室法检测其对细胞侵袭和迁移能力的影响。结果: MTT法检测DNP对6-10B细胞的非毒性浓度为0~10.0 μmol/L;8.0 μmol/L DNP处理6-10B细胞后,间接免疫荧光法检测发现AGR2蛋白表达明显上调,且以在细胞质中表达为主;不同浓度的DNP处理6-10B细胞24 h或用8.0 μmol/L的DNP处理6-10B细胞不同时间后,Western blot结果发现AGR2蛋白的表达水平增加且呈浓度和时间依赖性(P<0.05);siRNA-AGR2沉默6-10B细胞屮AGR2基因的表达后,DNP诱导6-10B细胞的侵袭及迁移能力较干扰前明显降低(P<0.05)。结论: DNP可能通过在转录水平上调AGR2的表达,影响鼻咽癌细胞的侵袭和转移能力。

关键词: 鼻咽癌, 二亚硝基哌嗪, AGR2, 肿瘤转移

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

中图分类号:

R73-37

王巍巍, 唐发清, 李跃进. 二亚硝基哌嗪上调AGR2促进鼻咽癌细胞转移[J]. 癌变·畸变·突变, 2016, 28(2): 115-120.

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.

参考文献

[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.

二亚硝基哌嗪上调AGR2促进鼻咽癌细胞转移

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	郭照萌, 张华, 张鹏. Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其机制[J]. 癌变·畸变·突变, 2021, 33(6): 420-425.
[2]	陈琪, 钟茜, 岳文涛. RNA结合基元蛋白24通过调控HOTAIR表达抑制鼻咽癌细胞增殖[J]. 癌变·畸变·突变, 2020, 32(4): 275-280.
[3]	黄蕾, 滕春莹, 雷鹏, 单毓娟. 伪足、有氧糖酵解与肿瘤细胞迁移的研究进展[J]. 癌变·畸变·突变, 2020, 32(1): 72-75.
[4]	贺荟静, 刘金成, 刘芡芡, 王云. 变异型EBER2通过抑制PKR通路增强鼻咽癌细胞抗凋亡能力[J]. 癌变·畸变·突变, 2018, 30(5): 333-338,344.
[5]	吕鹏, 满其忠, 王越, 庄正平, 王爱平, 曾益新. 抑制蛋白磷酸酶2A对鼻咽癌放射敏感性的影响[J]. 癌变·畸变·突变, 2016, 28(4): 262-268.
[6]	陈炯玉, 彭琳, 黄裔腾, 林雪琼. Prohibitin在放射照射诱导鼻咽癌细胞凋亡中的表达及亚细胞定位的变化[J]. 癌变·畸变·突变, 2016, 28(3): 200-204.
[7]	景绍武, 徐庆, 王军, 杨从容, 李广庆, 赵珍, 赵志军, 景尚华. E-钙黏蛋白异常表达与鼻咽癌临床病理关系的Meta分析[J]. 癌变·畸变·突变, 2016, 28(2): 97-102,106.
[8]	郑海霞, 崔永言, 申东兰, 彭安, 何艳玲. nm23-H1基因转染对肺癌L9981细胞黏附分子表达的影响[J]. 癌变·畸变·突变, 2015, 27(2): 134-136,141.
[9]	彭裕辉，谢长乐，庄珊珊，张利群，许镒洧. 联合检测p53自身抗体与EB病毒VCA-IgA对鼻咽癌的诊断价值[J]. 癌变·畸变·突变, 2015, 27(1): 26-29.
[10]	何迎春,刘宇勤,廖端芳,戴娜,何兰,范婧莹,田道法. 不同中医临床证型下行型鼻咽癌组织细胞核蛋白质差异表达的初步研究[J]. 癌变·畸变·突变, 2013, 25(3): 183-189.
[11]	叶静,李明华,张芳婷,万汇涓,龙霞. 5氮杂脱氧胞苷对鼻咽癌细胞RASSF1A基因甲基化和mRNA表达的影响[J]. , 2012, 24(3): 179-182.
[12]	杜泽森，吴智勇，郑春鹏，李卓毅，傅俊惠. 阴性淋巴结数目对胸段食管癌患者预后的影响[J]. , 2012, 24(1): 64-67.
[13]	李秀梅1,陈艳1,王洪江2,庞作良2,李卉1,姜孝芳1,古丽努尔·木哈依1,谌宏鸣1,李惠武1，*. 哈萨克族食管癌中MMP-1、MMP-2、MMP-7、TIMP-1 和MTA1的表达及其临床病理意义[J]. , 2011, 23(3): 190-193.
[14]	郑海霞1,周清华2,申东兰3,彭安3,何艳玲3,曹菊秀1. nm23-H1基因稳定转染肺癌细胞的鉴定[J]. , 2011, 23(2): 145-147.
[15]	孙　静;张　丹;朱庆均. 益气养阴方抑制肿瘤转移及对肿瘤相关基因表达谱的影响[J]. , 2009, 21(5): 348-352.