摘要
利用基因重组技术，将tpAcDNA与逆转录病毒载体LNSX重组构成LNS－ipA，转染病毒包装细胞，形成的重组逆转录病毒颗粒再用来感染牛血管内皮细胞。经G418筛选后的单克隆细胞培液，分别经纤维蛋白板法和发包底物法测取活性。结果：经基因转染的PA317细胞和牛血管内皮细胞，在含人纤维蛋白原和凝血酶的纤维蛋白板上出现溶圈；随培养时间的延长，细胞数量增加，tpA分泌活性也增加。结论：含人tpAcDNA转录单位的牛血管内皮细胞，其纤溶活性明显增高。该研究为将来进行基因治疗奠定了基础。
PURPOSE In an attempt to enhance the fibrinolytic activity of endothelial cells (EC) transduced with the human tissue - type plasminogen activator (tpA) gene.METHODS A recombinant retroviral vector containing tPA cDNA was constructed and transfected into bovine endothelial cells by retroviral vector - mediated gene transfer- The tpA activity of transduced cells was measeured with casein - plasminogen - agarose plate and a synthetic - peptide substract s - 239O.PESULTS The viral titer was 4 ×1O5 cfu/ml. The tpA activity of transduced cells was obviously increased. A greater number of transduced cells resulted in a higher level of tpA activity in the supernatant.CONCLUSIONS Retroviral vector - mediated gene transfer can be used to enhance the fibrinolytic activity of EC and the genetically engineered EC containing tpA cDNA might be applied to the prevention and treatment of thrombotic disease.
出处
《上海医科大学学报》
CSCD
1997年第6期450-452,共3页
Journal of Fudan University(Medical Science)
关键词
组织型
纤溶酶原激活剂
内皮细胞
血栓形成
tissue-type plasminogen activator
recombinant retrovirus
endothelial cell
transfection
