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热休克蛋白gp96在肝脏等多种肿瘤中过量表达,与肿瘤的恶性程度和患者不良预后呈显著相关性,其在肿瘤发生发展中作用机制有待深入探讨.通过生物信息学技术预测、荧光素酶报告基因检测、免疫印迹分析、实时定量PCR、RNA干扰,研究gp96 3′UTR作为ceRNA(competing endogenous RNA)对miR-642a和脱氧羟腐氨酸羟化酶(deoxyhypusine hydroxylase,DOHH)表达的影响.研究结果显示,miR-642a特异性靶向的野生型gp96 3′UTR,而不是miR-642a结合位点突变的gp96 3′UTR,可吸附下调miR-642a并同时上调miR-642a靶基因DOHH的表达,进一步研究发现gp96 3′UTR对DOHH的调控依赖于miR-642a.实验还发现DOHH并不影响gp96的表达.Gp96通过ceRNA上调DOHH的表达,为研究gp96促进肝癌等肿瘤的发生发展提供了新思路.
The overexpression of heat shock protein gp96 in many kinds of tumors such as the liver has a significant correlation with the degree of malignancy and the poor prognosis of patients and the mechanism of its action in tumorigenesis needs to be further explored.According to bioinformatics techniques, The expression of miR-642a and deoxyhypusine hydroxylase (DOHH) was examined by reporter gene assay, western blot analysis, real-time quantitative PCR and RNA interference. The results showed that the miR-642a-specific wild-type gp96 3’UTR, but not the gp96 3’UTR with miR-642a binding site mutation, could down-regulate the expression of miR-642a and simultaneously up-regulate the target gene DOHH of miR-642a Expression and further study found that gp96 3’UTR DOHH regulation dependent on miR-642a.Experiment also found that DOHH does not affect the expression of gp96.Gp96 through ceRNA up-regulation of DOHH expression for the study of gp96 promote the development of liver cancer and other provide New ideas.