应用端粒酶逆转录酶基因使猪血管内皮细胞永生化的初步研究

畜牧兽医学报 ›› 2007, Vol. 38 ›› Issue (4): 407-411.doi:

应用端粒酶逆转录酶基因使猪血管内皮细胞永生化的初步研究

苏正元;张彦明;洪海霞;刘建玲

1.西北农林科技大学动物科技学院，陕西杨凌 712100；2.西北大学生命科学学院，陕西西安 710069

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-04-25 发布日期:2007-04-25

Elementary Study on Immortalization of Porcine Vascular Endothelial Cell by Telomerase Reverse Transcriptase Transfection

Received:1900-01-01 Revised:1900-01-01 Online:2007-04-25 Published:2007-04-25

摘要/Abstract

摘要： 为寻求使猪主动脉血管内皮细胞增殖寿命延长的方法，本试验将含有的人类端粒酶催化亚基端粒酶逆转录酶(Human telomerase reverse transcriptase, hTERT)基因的真核表达质粒pCI-neo-hTERT通过脂质体介导法导入猪主动脉血管内皮细胞(porcine arterial endothelial cell , PAEC)。对转染后的血管内皮细胞进行Ⅷ因子和CD34鉴定，利用PCR-ELISA方法检测hTERT导入后细胞端粒酶活性表达情况，探讨了转染后细胞的生长特性。结果显示，转染后的细胞Ⅷ因子和CD34均呈阳性，证明转染后细胞仍表达血管内皮细胞的特异性蛋白；导入hTERT后, 细胞有较高的端粒酶活性, 增殖寿命较对照细胞延长了20代。表明hTERT导入细胞能重建细胞端粒酶活性, 从而延长细胞在体外培养的寿命。

Abstract: To elongate the proliferation life-span of porcine arterial endothelial cell(PAEC) in vitro. The eukaryotic expression plasmid pCI-neo-hTERT was transfected into PAEC by lipofection. FactorⅧ related antigen and CD34 of PAEC were identified after transfection by immunohistochemical method; we also detected telomerase activity of PAEC after transfection by PCR-ELISA. The results showed that Factor Ⅷ related antigen and CD34 in PAEC is positive after transfection. The PAEC after transfection expressed the vascular endothelial cell-specific protein still; Telomerase activity in PAEC is higher by hTERT introduction, the cell proliferation life-span was elongated to 20 population doublings compared to control. Conclusion, Telomerase activity can be activated in PAEC, thus the cell proliferation life-span was elongated in vitro.

苏正元;张彦明;洪海霞;刘建玲 . 应用端粒酶逆转录酶基因使猪血管内皮细胞永生化的初步研究[J]. 畜牧兽医学报, 2007, 38(4): 407-411.

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