反基因寡核苷酸抑制内皮细胞组织因子表达的体外研究

Effect of antigen oligonucleotide on inhibition of expression of the tissue factor in vitro

目的:探讨形成三股螺旋的寡核苷酸(triple helix-forming oligonucleotide,TFO)对内皮细胞组织因子(tissuefactor,TF)基因表达的抑制作用。方法:将针对TF基因启动子区的3个Spl/Egr-1位点合成的TFO,与含有人TF基因5’上游序列-244/+122 bp的荧光素酶报告基因质粒PL37,经脂质体转染内皮细胞,接受12 dyn/cm2切应力作用后,检测及分析报告基因活性。结果:反向硫代磷酸酯TFO(antiprarallel-phosphorothioated TFO,apTFO)显著抑制荧光素酶表达,抑制率高达70.3％。结论:apTFO能有效抑制切应力诱导的TF基因表达。

Objective: Tissue Factor (TF) is a cell membrane receptor protein that is the initiator of the extrinsic pathway of the blood coagulation cascade. Shear stress increases TF gene abnormally expression which plays a major role in thrombosis in the atherosclerosis vessel. This study was performed to investigate the affect of the triple helix-forming oligonucleotide (TFO) in inhibiting the expression of the endothelial tissue factor (TF). Methods: The endothelial celis of the human umbilical vein were transfected using DOTAP with the TFO constructed according to the three overlapping SPI/EGR - l sites and the luciferase report gene plasnid PL37 which containing the 5' upstream sequence -244/+122 bp of the human TF gene. Then the transfected celis were exposed to 12 dyn/cm2 of shear stress for 8h. Luciferase activity was detected. Resullts: The antiprarallel-phosphorothioated TFO (apTFO) can inhibit the expression of the TF gene by 70. 3%. Conclusion: The apTFO can efficiently inhibit the expression of the tissue factor induced by shear stress.