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Expression of Human Antithrombin Ⅲ Gene in Vascular Endothelial  like Cells

Objective To observe expression of human antithrombin Ⅲ (hAT-Ⅲ) gene in vascular endotheliallike cells(VELCs) after transfected. Methods Human bone marrow mesenchymal stem cells(BMMSCs) were isolated, cultured and proliferated in vitro, and were differentiated into VELCs. Then, the VELCs were divided into experimental group cells and control group cells randomly. Plasmid DNA with hAT-Ⅲ gene was transfected into VELCs by liposome mediate. At last, the hAT-Ⅲ expression was determined by reverse transcriptpolymerase chain reaction(RT-PCR), immunohistochemical stain(IHCS), Westernblotting and chromogenic substrate assay at 72h and 96h respectively. In the control group, the plasmid DNA was replaced by TE buffer, and the other methods were the same as the experimental group. Results RT-PCR showed that the specific DNA fragment of hAT-Ⅲ could be amplifed in the experimental group cells, none in the control group. IHCS showed positive expression of hAT-Ⅲ in the experimental group cells, negative in the control group. Westernblotting showed that the specific band of hAT-Ⅲ could be detected in the experimental group cells culture fluid, none in the control group. Chromogenic substrate assay showed that the hAT-Ⅲ activity of the experimental group cells was 9.50%±1.52%, the control group was 1.83%±1.17%， there was statistically difference between two groups(t=7.910，Plt;0.01). Conclusion The hAT-Ⅲ gene could be transfected into VELCs and expressed successfully.