重组逆转录病毒介导的人载脂蛋白AI基因在肌源细胞中的表达
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The Expression of Human Apolipoprotein Al in Myogenic Cells by Retrovirus Vector
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    摘要:

    探索一种安全有效的载脂蛋白AI基因表达系统。方法 构建含人载脂蛋白AI cDNA的双顺反子逆转录 病毒载体pLAEN.用其转化小鼠原代肌母细胞及肌源性细胞C2C12.ELISA法检测细胞液中人载脂蛋白AI的含量。结果 转化 后的小鼠原代肌母细胞及C2C12细胞在分化为肌管细胞后均获得异源表达人载脂蛋白AI的能力;而且经G418筛选也获得稳 定转化的C2C12细胞株,20天后仍保持有效表达人载脂蛋白AI的能力。结论 提示肌源性细胞在体外,以含人载脂蛋白AI cDNA 的逆转录病毒载体进行基田修饰后再移植回骨骼肌,有可能成为动脉粥样硬化基因治疗的一种可行方法。

    Abstract:

    To observe the expression of apolipoprotein A1 in transduced cultured muscle cells. Methods Bicistronic retrovirus vector containing human apolipoprotein A1 cDNA were constructed and packaged it into GP+E-86 and AM12 cell lines which was selected by G418. G418 resistant clone with high virus producing tiler was isolated. Mouse primary myoblasts and C2C12 mouse myoblasts were infected with recombinant virus supernatant. The content of apolipoprotein Al in cell culture medium was detected by ELISA. Results All of transduced mouse pti mary myoblasts and C2C12 cells gained and remained the ability for heterologous expression of human apolipoprotein Al even after differentiation into myotubes. Moreover, stable transduced C2C12 cell clones were generated by G418 selection, which effectively expressed human apolipoprotein Al up to 30 days. Conclusion The use of retrovirus vec tots to genetically modify myoblasts and then implantation back to skeletal muscle might be a safe and feasible strategy for gene therapy of atherosclerosis.

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魏恩会,陈琪,陈秀英,王南,范乐明,于书真.重组逆转录病毒介导的人载脂蛋白AI基因在肌源细胞中的表达[J].南京医科大学学报(自然科学版),2000,20(03):

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