PcDNA3.1/hTM质粒对真核细胞转染表达的实验研究

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

目的探讨含有人血栓调节蛋白(hTM)基因的真核表达质粒pcDNA 3.1/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pcDNA 3.1/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1 0%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pcDNA 3.1/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。

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 peDNA3. 1/hTM was 1.7 times higher than cells without transfeeted 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 transfeeted into endothelial cells by cationic liposome and the exogenous hTM protein had complete antieoagulation biological activity.