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中国癌症防治杂志

中国癌症防治杂志 ›› 2015, Vol. 7 ›› Issue (3): 173-177.doi: 10.3969/j.issn.1674-5671.2015.03.07

• 基础研究 • 上一篇    下一篇

二亚硝基哌嗪上调鼻咽癌细胞HSP70-2的表达及其意义

  

  1. 广东医学院附属佛山禅城中心医院检验科；暨南大学附属珠海医院/珠海市人民医院检验科
  • 出版日期:2015-06-25 发布日期:2015-07-13
  • 通讯作者: 唐发清
  • 基金资助:

    国家自然科学基金资助项目（81402368，81372282）

Dinitrosopiperazine-induced expression of heat shock protein 70-2 in nasopharyngeal carci-noma and its potential role in metastasis

  • Online:2015-06-25 Published:2015-07-13
  • Contact: tangfaqing33@hotmail.com

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摘要：

目的 分析二亚硝基哌嗪（dinitrosopiperazine，DNP）对鼻咽癌细胞热休克蛋白70-2（heat shock protein 70-2，HSP70-2）表达的影响，探讨DNP调控鼻咽癌细胞HSP70-2表达的意义和机制。方法 以HSP70-2低表达的低转移潜能鼻咽癌细胞6-10B为实验对象，用四甲基偶氮唑蓝（methyl thiazolyl tertrazolium，MTT）法检测DNP对6-10B细胞的非毒性浓度；分别采用细胞间接免疫荧光法、Western blot法和RT-PCR技术检测分析DNP对6-10B细胞HSP70-2表达的影响。结果 DNP对鼻咽癌6-10B细胞的非毒性浓度为0～100 μmol/L。经DNP处理6-10B细胞后，HSP70-2蛋白的表达显著增强，主要表达在细胞质。HSP70-2蛋白的表达与DNP处理的浓度呈剂量效应关系（P<0.05）。DNP（50 μmol/L、100 μmol/L）处理组HSP70-2 mRNA的表达水平与未处理组的表达无明显差异（P>0.05）。结论 DNP可能通过转录后调控机制上调鼻咽癌细胞HSP70-2蛋白的表达，推测DNP调控HSP70-2蛋白的表达并在鼻咽癌发生、发展中起重要作用。

关键词: 鼻咽肿瘤, 亚硝基化合物, 二亚硝基哌嗪, 热休克蛋白70-2

Abstract:

Objective To investigate dinitrosopiperazine（DNP）-induced expression of HSP70-2 in the nasopharyngeal carcinoma （NPC） cell line 6-10B，and examine whether it contributes to metastasis. Methods The MTT assay was used to screen for non-toxic concentrations of DNP，then the DNP was added to 6-10B cells to induce HSP70-2 expression. Protein levels were measured using indirect immunofluorescence and Western blotting. A dose-response curve for DNP-induced HSP70-2 expression was generated. HSP70-2 mRNA levels were quantitated using RT-PCR. The metastatic potential of 6-10B cells as a function of HSP70-2 expression level was determined. Results DNP at concentrations up to 100 μmol/L appeared to be non-toxic to 6-10B cells. Exposing these cells to DNP up-regulated HSP70-2 expression in a dose-dependent manner， with the protein localizing primarily to the cytoplasm. However， HSP70-2 mRNA levels were similar in treated and untreated cells. Conclusions DNP up-regulates HSP70-2 expression in 6-10B cells by post-transcriptional regulation，and this up-regulation may contribute to NPC metastasis.

Key words: Nasopharyngeal neoplasm, Nitroso-compound, Dinitrosopiperazine, Heat shock protein 70-2