维甲类X受体与维甲酸反应元件β_2RARE结合的研究

STUDY ON THE BINDING OF RETINOID X RECEPTOR WITH β 2 RARE

目的 :确定人真皮成纤维细胞中维甲类 X受体 ( RXR)与维甲酸反应元件 ( β2 RARE)的结合。方法 :培养成纤维细胞 ,反式维甲酸 ( At- RA)刺激细胞不同时段 ,抽提核蛋白 ,3 2 P标记β2 RARE作为探针 ,并加入 RXRα和β抗体 ,进行凝胶阻滞分析。结果 :对照组细胞核蛋白与β2 RARE形成复合体 ;At- RA刺激细胞后 ,迅速诱导产生明显的复合体 。选择两组核蛋白 (对照组和 At- RA刺激 4 h组 ) ,分别加入 RXRα和 β抗体 ,凝胶阻滞分析发现 RXRβ抗体可使这两组核蛋白产生新的上移复合体 SS,RXRα抗体则无此作用。结论 :RXRβ能与 β2 RARE结合 ,它们的结合不受 AT- RA的影响 ;提示 RXRβ可能是作为一辅助因子参与了 RARβ2

Objective:To confirm the binding of Retinoid X Receptors(RXR) with Retinoic Acid Responsive Element (β 2RARE) Method:Fibroblasts in human derma were cultured with stimulation for All trans Retinoic Acid (At RA) Labeling β 2RARE with 32 P as a probe,nuclear extract was isolated from these cells and incubated with antibody to RXR α and β,then gel retardation assays were performed Results:The nuclear protein from control group was binded with β 2 RARE to form a complex Ⅰ Stimulating cells with At RA quickly induced an obvious complex Ⅱ Two groups of nuclear protein (control group and the group of 4 h stimulated with At RA) were selected for incubating separately with antibody to RXR α and β The gel retardation assays showed a new supershift complex SS after using antibody to RXR β for incubating with the nuclear proteins of these two groups,while the antibody to RXR α did not appear to have this effect Conclusion:RXR β binded to β 2 RARE AT RA has no effect on this binding It suggested that RXR β may be an accessory factor in transcriptional regulation of RARβ 2