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0～100μmol/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。
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.
出处
《中国癌症防治杂志》
CAS
2015年第3期173-177,共5页
CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT
基金
国家自然科学基金资助项目(81402368
81372282)
