促血小板生成素基因转染小鼠骨髓基质细胞体外表达的实验研究

Expression of thrombopoietin in vitro after transfected into mouse bone marrow stromal cells

目的探讨促血小板生成素(thrombopoietin,TPO)基因修饰的小鼠骨髓基质细胞(bone marrow stromal cells,BMSCs)体外表达TPO的变化特点。方法以脂质体介导法将TPO cDNA真核表达质粒转染BMSCs,RT-PCR方法检测TPO mRNA的表达,免疫组化法及Western blot测定TPO蛋白的表达。结果转染TPO基因的BMSCs其TPO mRNA及TPO蛋白表达量明显高于空载体组(P<0.01)及未转染组(P<0.01)。结论阳离子脂质体能有效介导TPO cDNA真核表达质粒转染BMSCs,且转染后BMSCs中TPO mRNA及TPO蛋白的表达显著上调。

Objective To investigate the changes of thrombopoietin （TPO） expression in mouse bone marrow stromal cells （BMSCs） in vitro after TPO gene transfection. Methods The bone marrow stromal cells of Balb/C mice aged 6 to 8 weeks were isolated and randomly divided into control group, pcDNA3.1 vector group and TPO transfection group. After TPO cDNA eukaryotic expressing vector encapsulated lipofectamineTM 2000 was transfected into BMSCs for 72 h, the expression of TPO mRNA and TPO protein in BMSCs were detected by RT-PCR and immunohistochemistry, Western blotting. Results The expressions of TPO mRNA and TPO protein in TPO gene transfected BMSCs were significantly enhanced as compared to pcDNA3. 1 vector group and normal control BMSCs （ all P 〈 0.01 ）. Conclusion TPO cDNA eukaryotic expression vector could be transfected into BMSCs efficiently by lipofectamineTM 2000, and the expressions of TPO mRNA and TPO protein were up-regulated in TPO gene transfected BMSCs.