凝血栓蛋白-1Ⅰ型重复序列反义核酸在血管内皮细胞中的作用研究

目的探讨TSP-1Ⅰ型重复序列反义RNA对人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HU-VECs)生长活性、增殖活性影响。方法构建TSP-1Ⅰ型重复序列反义RNA真核表达载体(pcDNA3.1-/anti-TSP-1-Ⅰ),经双酶切、测序及Western blot鉴定后,转染HUVECs。采用MTT法检测转染后HUVECs活性改变,流式细胞仪检测转染后HUVECs周期变化,透射电镜观察转染后HUVECs形态学改变。结果构建的pcDNA3.1-/anti-TSP-1-Ⅰ经鉴定后转染HUVECs,MTT法检测所得HUVECs OD值较未转染组、pcDNA3.1-空载体转染组升高(均P<0.01);流式细胞仪检测结果显示:pcDNA3.1-/anti-TSP-1-Ⅰ转染HUVECs后S+G2(%)较未转染组、pcDNA3.1-空载体转染组S+G2(%)延长(均P<0.01);透射电镜结果显示:pcDNA3.1-/anti-TSP-1-Ⅰ转染后HUVECs核仁相对增多。结论TSP-1反义RNA能够促进HUVECs增殖和生长。

Effects of thrombospondin-1 typeⅠ repeat antisense RNA on human endothelial cells

Aim To investigate the effects of thrombospondin-1 type Ⅰ repeat antisense RNA on cell proliferation and growth activity of human endothelial cells.Methods The eukaryotic expression vector of pcDNA3.1-/anti-TSP-1-Ⅰ was constructed.Then the vector was transfected into human umbilical vein endothelial cells(HUVECs).The growth activity of endothelial cells after transfection was detected by MTT.The variation of endothelial cell's cycle was determined by flow cytometry.Cell morphological change was observed by transmission electron microscope.Results The eukaryotic expression vector(pcDNA3.1-/anti-TSP-1-Ⅰ)was successfully constructed and transfected into HUVECs.The OD value of pcDNA3.1-/anti-TSP-1-Ⅰ transfective group was promoted compared with that of nontransfective group(P<0.01)and pcDNA3.1-vector group(P<0.01);The S+G(%)of pcDNA3.1-/anti-TSP-1-Ⅰtransfective group was abridged compared with that of nontransfective group(P<0.01)and pcDNA3.1-vector group(P<0.01).The nucleole increased in the group of pcDNA3.1-/anti-TSP-1-Ⅰtransfective group.Conclusion Thrombospondin-1 type Ⅰ repeat antisense RNA can accelerate proliferation and growth of human endothelial cells.