脂质体介导pcDNA3.1-IDO转染hepG_2细胞的初步研究

Preliminary study of human hepatoma carcinoma cell line hepG2 transfected with plasmid pcDNA3.1-IDO by lipofectamine

目的:建立脂质体介导pcDNA3.1-IDO转染人肝癌细胞株hepG2细胞的转染方法。方法:在大肠杆菌中扩增pcDNA3.1-IDO质粒,通过lipofectamineTM2000转染试剂将pcDNA3.1-IDO转入人肝癌细胞hepG2,同时设置空质粒转染组(pcDNA3.1-hepG2细胞)和空白对照组(hepG2细胞)。分别应用RT-PCR和Western blot方法检测吲哚胺2,3-双加氧酶(IDO)基因及IDO蛋白在hepG2细胞中的表达。结果:经RT-PCR和Western blot检测证实空质粒转染组和空白对照组hepG2细胞无IDO基因及蛋白的表达,而重组质粒组的hepG2细胞均表达IDO基因和蛋白。结论:通过脂质体介导成功地将pcDNA3.1-IDO转染入肝癌细胞株hepG2细胞。这为研究IDO基因奠定了基础。

OBJECTIVE：To determine the transfection expression of plasmid pcDNA3.1-IDO in human hepatoma carcinoma cell line hepG2,and to establish a transfection method of hepG2.METHODS：The plasmid pcDNA3.1-IDO was amplified in Escherichia coli.The cultured hepG2 cells were transfected with pcDNA3.1-IDO by lipofectamineTM2000 reagent.The hepG2 cells and hepG2 cells trancfected with blank plasmid pcDNA3.1（pcDNA3.1-hepG2 cell）were used as control group.The transient expression of IDO in hepG2 cells transfected with recombinant plasmid was determined by RT-PCR and Western blot.RESULTS：IDO gene and protein were expressed transiently in hepG2 cells transfected with recombinant plasmid shown by RT-PCR and Western blot,respectively.CONCLUSION：The IDO gene was successfully transfected into human hepatoma carcinoma cell line hepG2 by means of lipofectamineTM2000 reagent,which provides the basis for further studies of IDO gene.