Construction of Expression Vector pcDNA3.1(+)TPA and Study of Its Expression Activity in Human Umbilical Vein Endotheliocytes / 中国医师杂志

ABSTRACT

Objective To establish the expression of exogenous tissue plasminogen activator (TPA) gene in human umbilical vein endotheliocytes to provide both theoretical basis and new method for gene therapy of ischemic heart disease and prevention of postoperative vessel re-stenosis. Methods Expression vector pcDNA3 1(+)TPA was constructed, and transfected into human umbilical vein endotheliocytes cultivated in vitro by lyposome. The exogenous TPA expression was observed. Results The expression vector pcDNA3 1(+)TPA could efficaciously express TPA in human umbilical vein endotheliocytes. The protein quantity of TPA in the transfected cells was 568 6ng/10 6cell/24h, when measured by enzyme-linked immunosorbent assay, while that in the non-transfected cells was 17 8ng/10 6 cell/24h. The exogenous TPA's activity in the transfected cells was 108 8IU/10 6cell/24h, when measured by chromogenic substrate assay, while that in the non-transfected cells was 5 6 IU/10 6cell/24h. Conclusion When pcDNA3 1(+)TPA was transfected into human umbilical vein endotheliocytes, exogenous TPA could be expressed efficaciously, which provides theoretical basis and new method for TPA gene therapy of ischemic heart disease.