首页> 中文期刊> 《中国癌症防治杂志》 >二亚硝基哌嗪上调鼻咽癌细胞HSP70鄄2的表达及其意义

二亚硝基哌嗪上调鼻咽癌细胞HSP70鄄2的表达及其意义

         

摘要

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 100μmol/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.%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0~100μmol/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(50μmol/L、100μmol/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。

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