PcDNA3.1/hTM质粒对真核细胞转染表达的实验研究

目的： 探讨含有人血栓调节蛋白（hTM）基因的真核表达质粒pcDNA3.1/hTM转染脐静脉内皮细胞（HUVECs）的抗凝效应。方法：由阳离子脂质体介导将pcDNA3.1/hTM质粒转入内皮细胞中，以RT-PCR法检测hTMmRNA的表达;免疫组化法检测hTM分子的表达。结果：转染重组质粒的HUVECs表达的hTMmRNA约为载体质粒组及未转染组的1.7倍。免疫组化显示有平均10%的HUVECs获得转染，阳性细胞较阴性细胞hTM的表达强度有明显提高。结论：pcDNA3.1/hTM质粒能被导入 HUVECs中表达，而且外源性hTM分子具有完全的抗凝生物学活性。

A study of endothelial cells modified by pcDNA3.1/hTM

Objective To identify the anticoagulation effect of pcDNA3.1 transfected exogenous human (throbomodulin) molecule(hTM) on umbilical vein endothelial cells(HUVECs).Methods The pcDNA(3.1)/hTM plasmid was transduced into HUVECs by cationic liposome.Expression of hTMmRNA and hTM(protein) was detected by RT-PCR and immunohistochemistry,respectively.Results The expression of(hTMmRNA) in HUVECs transduced with pcDNA3.1/hTM was 1.7 times higher than cells without transfected PcDNA3.1/hTM.Immunogistochemical examination showed about 10% cell tranfected with PcDNA3.1/hTM and high-level hTM protein was detected on positive cells.Conclusions The pcDNA3.1/hTM plasmid could be transfected into endothelial cells by cationic liposome and the exogenous hTM protein had complete anticoagulation biological activity.