二亚硝基哌嗪上调HSP70-2促进鼻咽癌细胞转移

发布时间：2018-08-16 17:32

【摘要】：目的:探讨二亚硝基哌嗪(dinitrosopiperazine,DNP)通过调控热休克蛋白70-2(heat shock protein70-2,HSP70-2)表达参与鼻咽癌转移的分子作用机制。方法:以具有低转移潜能的鼻咽癌细胞6-10B作为研究材料,应用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、蛋白质印迹法和实时荧光定量-PCR法检测DNP对6-10B细胞中HSP70-2表达的影响;采用特异性针对HSP70-2基因的小干扰RNA(small interfering RNA,siRNA)-HSP70-2沉默HSP70-2的表达,并用Transwell小室法检测DNP对细胞侵袭和迁移能力的影响。结果:DNP对6-10B细胞的非毒性浓度为0~100μmol/L:DNP(100μmol/L)处理6-10B细胞后,HSP70-2蛋白表达明显上调,且以在细胞质中表达为主;不同浓度DNP(50和100μmol/L)处理6-10B细胞后,HSP70-2蛋白的表达水平呈浓度依赖性(P0.05);DNP(50和100μmol/L)处理组中HSP70-2 mRNA的相对表达量分别为0.81±0.14和0.81±0.26,同空白对照组的1.04±0.33比较,差异无统计学意义(P0.05)。siRNA-HSP70-2沉默6-10B细胞中HSP70-2蛋白的表达可抑制DNP诱导的6-10B细胞的侵袭及迁移能力。结论:DNP可能通过转录后调控机制上调HSP70-2表达,参与鼻咽癌细胞的侵袭和转移。
[Abstract]:Aim: to investigate the molecular mechanism of dinitro-opiperazine (DNPs) involved in the metastasis of nasopharyngeal carcinoma (NPC) by regulating the expression of heat shock protein 70-2 (HSP70-2). Methods: the nontoxic concentration of DNP on 6-10B cells was detected by MTT assay, and the indirect immunofluorescence assay was used to detect the nontoxic concentration of 6-10B cells with low metastatic potential. The effect of DNP on the expression of HSP70-2 in 6-10B cells was detected by Western blot and real-time fluorescence quantification-PCR, the expression of HSP70-2 was silenced by small interfering RNA (small interfering RNA-siRNA targeting HSP70-2 gene) -HSP70-2, and the effect of DNP on the invasion and migration of 6-10B cells was detected by Transwell chamber assay. Results the expression of HSP70-2 protein was up-regulated in 6-10B cells treated with 0 渭 mol/L:DNP (100 渭 mol/L) of 0 渭 mol/L, and the expression of HSP70-2 protein was mainly in the cytoplasm of 6-10B cells. The expression level of HSP70-2 protein in 6-10B cells treated with different concentrations of DNP (50 渭 mol/L and 100 渭 mol/L) was in a dose-dependent manner (P0.05). The relative expression of HSP70-2 mRNA was 0.81 卤0.14 and 0.81 卤0.26 in the treatment group of 50 渭 mol/L and 100 渭 mol/L, respectively, compared with 1.04 卤0.33 in the blank control group. There was no significant difference (P0.05). SiRNA-HSP70-2 silenced the expression of HSP70-2 protein in 6-10B cells, which inhibited the invasion and migration of 6-10B cells induced by DNP. Conclusion the expression of HSP70-2 in nasopharyngeal carcinoma cells may be involved in the invasion and metastasis of nasopharyngeal carcinoma cells through the mechanism of posttranscriptional regulation.
【作者单位】：暨南大学附属珠海医院;珠海市人民医院检验科;中南大学湘雅医院检验科;
【基金】：国家自然科学基金资助项目(编号:30973400,81071718,81372282) 教育部新世纪优秀人才支持计划(编号:NCET-06-0685)
【分类号】：R739.63

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