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目的探讨HMGB1在外源DNA-poly(dA-dT)刺激细胞Ⅰ型干扰素IFN-β中的作用。方法荧光素酶分析方法检测经poly(dA-dT)刺激后细胞IFN-β的表达差异,同时PCR法检测了胞内重要DNA结合蛋白—高迁移率族蛋白-1(HMGB1)的表达改变;构建携带HMGB1特异性shRNA的慢病毒载体,获得HMGB1稳定下调(HMGB1KD)细胞,经poly(dA-dT)刺激后检测IFN-β表达,分析HMGB1在外源DNA刺激IFN-β产生中的作用。结果 poly(dA-dT)刺激293T细胞后,IFN-β表达显著上调,同时,HMGB1的表达在不同细胞中也明显上升,当下调HMGB1后,poly(dA-dT)刺激IFN-β产生的能力大幅削弱,表达水平降至对照组的1/8(P<0.05)。结论 HMGB1在促进外源DNA诱导Ⅰ型干扰素产生中发挥了关键作用。
Objective To investigate the role of HMGB1 in exogenous DNA-poly (dA-dT) -stimulated type I IFN-β. Methods Luciferase assay was used to detect the expression of IFN-β in cells after stimulation with poly (dA-dT). The expression of HMGB1, an important intracellular DNA binding protein, was detected by PCR. The lentiviral vector carrying HMGB1 shRNA was constructed and HMGB1 stably down-regulated (HMGB1KD) cells were obtained. The expression of IFN-β was detected by poly (dA-dT) stimulation and the effect of HMGB1 on IFN-β production by exogenous DNA was analyzed. Results After poly (dA-dT) stimulated 293T cells, the expression of IFN-β was significantly up-regulated while the expression of HMGB1 was also significantly increased in different cells. The ability of poly (dA-dT) to stimulate IFN- Significantly weakened, and the expression level dropped to 1/8 of the control group (P <0.05). Conclusion HMGB1 plays a key role in promoting the production of type Ⅰ interferon induced by exogenous DNA.