转染bcl-2基因对心肌细胞缺氧耐受性的影响

Effect of transfecting bcl-2 gene into cardiomyocytes on their hypoxic tolerance in vitro

目的 探讨转染bcl 2基因能否提高心肌细胞对缺氧的耐受性。方法 ①重组真核细胞表达载体 :将含编码人bcl 2cDNA完整开放阅读框的bcl 2cDNA重组至逆转录病毒载体 pLXSN的EcoRⅠ位点上 ,用脂质体包裹重组质粒导入PA317细胞株 ,并经G4 18筛选阳性克隆 ;②用含高滴度重组病毒的上清感染心肌细胞 ,原位杂交鉴定其转染是否成功 ;③观察转染心肌细胞在缺氧环境下的生存率。结果 ①重组了含bcl 2的真核细胞表达质粒 ;②转染bcl 2基因能明显提高心肌细胞对缺氧的耐受能力。结论 转染bcl 2基因能显著提高心肌细胞在缺氧环境下的存活能力。

Objective To study whether transfecting bcl 2 gene into cardiomyocytes can improve their hypoxic tolerance in vitro. Methods bcl 2 cDNA containing human full length open reading frame was recombined into EcoR I site of retrovirus vector pLXSN. The plasmids were packed with liposome and transfected into cell line PA317. PA317/s bcl 2 was selected from the transfected cells by its resistance to G418. Cardiomyocytes were infected with high titer recombined virus produced by PA317/s bcl 2. The transfection was identified with in situ hybridization. The growth properties and tolerance to hypoxia of the cardiomyocytes were studied in bcl 2 gene transfected cardiac and non cardiac cells. Results Eukaryotic cell expression plasmid containing bcl 2 gene was constructed and it could increase the survival of cardiomyocytes under hypoxic condition. Conclusion Transfecting bcl 2 gene into cardiacmyocytes can significantly enhance their hypoxic survival rate.