| IDO基因稳定转染HepG2细胞系的建立 |
| 投稿时间：2011-07-12 修订日期：2011-07-27 点此下载全文 |
| 引用本文：刘春亮,冯惠枝,刘燕,卜晓倩,申慧琴,张瑞,唐运萍,王琦.IDO基因稳定转染HepG2细胞系的建立[J].医学研究杂志,2012,41(2):68-70 |
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| 基金项目:山西省科研攻关项目（200903110） |
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| 中文摘要:目的建立稳定转染IDO基因的HepG2细胞系，为进一步研究IDO基因在肝癌免疫逃逸中的作用奠定基础。方法应用脂质体将真核细胞表达质粒pcDNA3.1-IDO和空载质粒pcDNA3.1转染入HepG2细胞，经G418进行筛选后，挑取单克隆进行培养，用反转录酶聚合酶链反应(RT-PCR) 和Western blot方法验证获得的表达细胞株。结果经RT-PCR和Western blotting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细胞表达IDO基因和蛋白。结论pcDNA3.1-IDO质粒体外稳定转染人肝癌细胞HepG2，可以有效地使IDO基因在RNA水平和蛋白水平均表达，成功建立了稳定转染基因IDO的HepG2细胞系，为进一步探讨该基因的功能奠定了一定基础。 |
| 中文关键词:吲哚胺2，3-双加氧酶（IDO） HepG2细胞 稳定转染 |
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| Construction of a HepG2 Cell Line with IDO Gene Steady Transfection |
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| Abstract:ObjectiveTo establish a human hepatoma carcinoma cell line with IDO gene steady transfection and verify the gene expression of IDO in HepG2 cell, which provides the foundation for the tumorous immune escape effect of pcDNA3.1-IDO.MethodsThe eukaryotic expression plasmid pcDNA3.1-IDO and empty vector pcDNA3.1 were transfected into HepG2 cells with lipofectine.G418 was used for selecting the positive clone.After being screened with G418,the single cell clone was sought out and cultured.The expression of IDO in HepG2 cells was detected by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting. ResultsNot only at the level of transcription but also translation, IDO gene expressed in IDO transfectant confirmed by RT-PCR and Western blotting respectively. ConclusionStabily transfected HepG2 of pcDNA3.1-IDO Plasmid can lead to upregulating IDO mRNA and protein expression obviously,which showes transfection HepG2 of pcDNA3.1-IDO successfully. |
| keywords:Indoleamine 2,3-dioxygenase(IDO) HepG2 cell Steady transfection |
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