雷洛昔芬对血管内皮细胞ERE转录活性的影响及与雌激素受体亚型的关系

Effects of raloxifene on the ERE transcription activity and its relationship with estrogen receptor subtypes in vascular endothelial cells

目的:研究雷洛昔芬对血管内皮细胞雌激素应答元件(ERE)转录活性的影响与雌激素受体(ER)亚型的关系。方法:采用腺病毒载体将雌激素受体α型(ERα)或雌激素受体β型(ERβ)导入培养的血管内皮细胞,构建ERE荧光素酶报告基因质粒ERE鄄TK鄄Luc,采用真核细胞转染技术将ERE鄄TK鄄Luc与内参照质粒PRL鄄SV40共同转染血管内皮细胞,双荧光素酶报告法检测雷洛昔芬刺激的转染细胞荧光素酶的相对活性。结果:导入ERα细胞,雷洛昔芬刺激后荧光素酶活性为0.0247±0.0008,显著低于未接受雷洛昔芬刺激细胞的0.0418±0.0007穴P<0.001雪;导入ERβ的细胞,雷洛昔芬刺激后荧光素酶活性为0.0304±0.001,未接受雷洛昔芬刺激的细胞为0.0306±0.003,二者相比无明显变化(P>0.05)。结论:雷洛昔芬显著抑制ERα介导的血管内皮细胞ERE转录活性。

Objective: To investigate the effect of Raloxifene on the estrogen responsive element (ERE) transcription activity and its relationship with estrogen receptor subtypes in vascular endothelial cells. Methods: ERE-TK-Luc, which contains a firefly luciferase reporter vector, was used as a reporter plasmid. Vascular endothelial cells were transfected with ERE-TK-Luc and control plasmid PRL-SV40 via FuGENE 6. Two kinds of luciferase activity were measured by dual-luciferase reporter assay system. The luciferase activity in transfected cells treated with Raloxifene (10-7mol/L) was compared with that in untreated cells. Replication-deficient adenovirus vectors carrying the coding region of human ERα or ERβ were constructed and transferred into vascular endothelial cells, and the luciferase activities of raloxifene-treated and -untreated cells were measured and compared. Results: The luciferase activity in cells treated with Raloxifene decreased compared with that in untreated cells (P<0.05, n=3). In cells transferred with ERα, the luciferase activity decreased significantly (P<0.001, n=3). But in cells transferred with ERβ, no significant change was found. Conclusion: Raloxifene inhibits the ERE transcription activity induced by ERα in vascular endothelial cells.