反应元件CRE和SRE参与猪CCR1和CCR5信号通路的分析

Analysis of CRE and SRE Response Element in the Porcine CCR1 and CCR5 Signal Pathway

为探明调节活化正常T细胞表达与分泌的趋化因子(RANTES)与趋化因子受体1(CCR1)、趋化因子受体5(CCR5)的信号通路关系,采用分子克隆技术、Western blotting技术构建、鉴定猪CCR1和CCR5真核表达载体,通过双荧光素酶报告系统检测RANTES分别作用CCR1和CCR5后萤火虫荧光素酶的表达量间接反映出cAMP反应元件(CRE)和血清反应元件(SRE)的表达水平。结果显示,RANTES作用于CCR1能降低HEK293T细胞内CRE的表达水平,对SRE有激活作用,而作用于CCR5后既能提高CRE的表达水平,也能增强SRE的表达。结果证明,RANTES通过CCR1受体能激活SRE,降低由毛喉素升高的CRE表达水平;RANTES通过CCR5受体对CRE和SRE均有激活作用。

The aim of the present study was to explore the signaling relationship between the chemokine regulated on activation,normal T cell expressed and secreted(RANTES)and its receptors[chemokine receptor 1(CCR1)and chemokine receptor 5(CCR5)].The molecular cloning techniques and Western blotting were used to construct eukaryotic expression vectors,and then used dual luciferase report system tests to detect the expression of firefly luciferase after receptors activated or not with RANTES,to indirectly reflect the amount of cAMP response element(CRE)and serum response element(SRE)expression level.The results showed that CCR1 activated with RANTES could reduce the level of CRE expression and enhance the level of SRE expression in HEK293 Tcell,and CCR5 activated with RANTES could enhance the level of CRE expression and SRE expression.The results proved RANTES interacted with CCR1 could stimulate the level of SRE expression and reduce the level of CRE expression increased by Forskolin,with CCR5 could enhance the expression level of CRE and SRE.