中文摘要：

目的：探讨二亚硝基哌嗪（dinitrosopiperazine，DNP）通过调控热休克蛋白70—2（heatshockprotein70—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-CR法检测DNP对6-10B细胞中HSP70—2表达的影响；采用特异性针对H15P70—2基因的小干扰RNA（smallinterferingRNA，siRNA）一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0～100gmol／L；DNP（100gmol／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100gmol／L）处理6-10B细胞后，HSP70—2蛋白的表达水平呈浓度依赖性（P〈0．05）；DNP（50和100gmol／L）处理组中HSP70—2mRNA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（P〉0．05）。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。

英文摘要：

Objective： To investigate the mechanism of dinitrosopiperazine （DNP）-mediated heat- shock protein 70-2 （HSP70-2） expression involved in the metastasis of nasopharyngeal cancer （NPC）. Methods： Non-cytotoxic concentration of DNP against NPC cell line 6-10B with low metastatic potential was determined by MTT assay. The expressions of HSP70-2 protein and mRNA in 6-10B cells treated with DNP （50 and 100 μmol/L） were detected by indirect immunofluorescence assay, Western blotting, and real-time fluorogenic quantitative-PCR, respectively. The expression of HSP70-2 was silenced by small interfering RNA （siRNA）-HSP70-2. The capabilities of invasion and migration of 6-10B cells treated with 1 00 μmol/L DNP were detected by Transwell assay. Results： The non-cytotoxic concentration of DNP against 6-10B cells was 0-100 μmol/L. After treatment with 100 μmol/L DNP, the expression of HSP70-2 was significantly increased and mainly located in the cytoplasm. This expression was also in a dose- dependent manner. The relative expression levels of HSP70-2 mRNA in 6-10B cells treated with 50 and 100 μmol/L DNP were 0.81±0.14 and 0.81±0.26, respectively, which were not significantly different from that of the blank control group （vs 1.04±0.33, P 〉 0.05）. The capabilities of invasion and migration in 6-10B cells were inhibited by silencing the expression of HSP70-2 induced by siRNA-HSP70-2. Conclusion： DNP can up-regulate the expression of HSP70-2 in 6-10B cells through post-transcriptional regulation mechanism and then mediate the metastasis of NPC cells.