前期研究发现DNP通过HSP70-2促进鼻咽癌转移，HSP70-2APTase活性显著增高，分子模拟预测DNP与HSP70-2-J-domain结合。由此推测DNP与HSP70-2-J-domain结合，增强HSP70-2APTase活性参与鼻咽癌转移。本项目拟研究：①分析DNP浓度、HSP70-2APTase活化与鼻咽癌转移关系；②构建HSP70-2-J-domain质粒，转染Hsp70-2-/-细胞，分析3H-DNP与J-domain结合、HSP70-2APTase活化和细胞转移；③构建HSP70-2-J-domain突变体，分析DNP不与J-domain结合对HSP70-2APTase活性和细胞转移的影响。④将HSP70-2及突变体导入Hsp70-2-/-细胞，接种至裸鼠，动物实验分析细胞转移。阐明DNP通过活化HSP70-2APTase参与鼻咽癌转移的机理，为鼻咽癌防治提供科学依据

Our previous works found that N,N'- dinitrosopiperazine (DNP) mediates nasopharyngeal carcinoma (NPC) through HSP70-2, following an increased HSP70-2APTase activity. Molecular dynamics simulations data showed that DNP binds to HSP70-2-J-domain. Based on these, we speculate that DNP may increase HSP70-2APTase activity through binding to HSP70-2-J-domain, and promotes NPC metastasis. In this project, we want to ①analyze the relationship of DNP concentration, HSP70-2APTase activity and NPC metastasis, and confirm whether DNP-mediated HSP70-2APTase activity involves NPC metastasis; ②construct HSP70-2-J-domain plasmids, transfect them to Hsp70-2-/- cells, analyze the interaction of 3H-DNP and J-domain, HSP70-2APTase activity and cell metastasis; ③construct HSP70-2-J-domain mutant dominant plasmids, transfect the plasmid to NPC cells, analyze DNP-mediated NPC metastasis when DNP dose not interact with J-domain; ④transfect HSP70-2 and its mutant dominant plasmids into Hsp70-2-/- cell, and the stable cell lines containing J-domain, HSP70 mutant dominant were obtained, these stable cell line were injected into nude mice, and DNP-mediated NPC metastasis though binding to J-domain-HSP70-2 were confirmed using animal experiments. The mechanism of DNP-mediated NPC metastasis though activating HSP70-2-APTase will be clarified, these will provide a novel clue for NPC prevention and therapy.