canstatin基因在体外培养血管内皮细胞中的表达及其意义

Expression of Canstatin Gene in the HUV-ECC Cells in vitro and Its Meaning

目的探讨canstatin基因通过电穿孔法在体外培养人脐静脉内皮细胞中的表达及对其生长与凋亡的影响。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RT-PCR检测canstatin在转基因细胞mRNA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。结果检测到canstatin在转染人脐静脉内皮细胞中的表达,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。

Objective To explore the effects of canstatin gene transfection by electroporation on the growth and apotosis of HUV-ECC cells 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. Expression of canstatin mRNA in the transfected cells was examined by RT-PCR assay, and the cell cycles were analysed by flow cytometry. Results The expression of canstatin mRNA was found in the transfected HUV-ECC cells. The percentage of cell apoptosis in the transfected HUV-ECC group（ 16. 90% ） was significantly increased compared with those in the control group（ 1.47% ） and the parental cell group（2.85% ）（ P 〈0.01 ）. The growth of the transfected cells was significantly inhibited. Conclusions The canstatin gene can be expressed in the HUV-ECC cell line after the transfection. Canstatin can inhibit endothelial cells proliferation and induce their apoptosis.