Canstatin基因转染对体外培养血管内皮细胞增殖与凋亡的影响

Effects of Canstatin gene transfection on growth and apotosis of HUV-ECC cells in vitro

目的探讨电转染Canstatin基因对体外培养人脐静脉内皮细胞生长的影响。方法将Canstatin基因通过电穿孔法转染人脐静脉内皮细胞HUV-ECC,行G418筛选获得转基因细胞克隆。用SDS-PAGE电泳检测Canstatin蛋白在转基因细胞培养上清液中的表达,以流式细胞仪分析细胞周期,并比较转基因和未转基因细胞的生长特性。结果Canstatin在转染人脐静脉内皮细胞中表达并分泌至上清液中,Canstatin基因转染内皮细胞组的凋亡率(16.90%)高于空载体组(1.47%)和亲代细胞组(2.85%,P<0.01),转染细胞生长明显受抑。结论Canstatin能特异地抑制血管内皮细胞增殖,并诱导细胞凋亡。

AIM To explore the effects of Canstatin gene transfection by electroporation on the growth of HUV-ECC cell in vitro. METHODS The expression vector of pCMV-Script/Canstatin was transfected into HUV-ECC cells by electroporation and the positive clone was screened by G418. The growth characteristics of cell line before and after transfection were compared. Canstatin protein in supernatant was examined by SDS-PAGE asssy and the cell cycles were analyzed by flow cytometry. RESULTS The expression of Canstatin gene at the level of protein was found in supernatant of the transfected HUV-ECC cells. The percentage of apoptosis of the transfected HUV-ECC cells （ 16.90% ） was markedly increased compared with that of the control group （1.47%） and the parental cell group （2.85% ,P 〈0.01 ）. The growth of the transfected cells was significantly inhibited. CONCLUSION The Canstatin gene is expressed in the HUV-ECC cell line after transfection and the Canstatin protein can inhibit endothelial cell＇s proliferation and induce its apoptosis.