靶向性重组PF4裸质粒抗血管生成活性的研究

Chemical Industry and Engineering（2017）

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Abstract

构建重组真核表达载体pcDNA3.1(+)-PF447-70-RGD,并检测靶向性重组血小板因子四(PF4)在真核细胞内的表达水平,探讨其体外抗血管生成活性.设计构建了靶向性重组PF4真核表达载体pcDNA3.1(+)-PF447-70-RGD,用脂质体法将其转染至中华仓鼠卵巢细胞系(CHO),用G418筛选稳系,采用RT-PCR和Western blot检测表达产物,MTT法测定CHO稳转体系培养土清对脐静脉内皮细胞(HUVEC)增殖的影响.成功构建出重组真核表达载体pcDNA3.1(+)-PF447-70-RGD,并获得稳转CHO细胞系,从基因水平和蛋白水平均可检测到目的基因的表达.MTT法显示,CHO稳系培养上清对HUVEC生长抑制率是50.8％,与对照组比,血管内皮细胞的增殖能力显著减小.靶向性重组PF4具有较强的体外抑制血管生成活性,该重组肽能够抑制HUVEC增殖.

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