中国组织工程研究Issue(1)：57-62,6.DOI:10.3969/j.issn.2095-4344.2014.01.010
低氧诱导因子1α慢病毒载体转染骨髓间充质干细胞的成骨基因表达
Influence of hypoxia-inducible factor-1 alpha lentiviral vector on osteogenic gene expression of rabbit bone marrow mesenchymal stem cells
摘要
Abstract
BACKGROUND:Osteogenesis is closely integrated with angiogenesis in bone formation and repair process, and hypoxia-inducible factor 1 alpha (HIF-1α) is considered to be the most important core transcription factor promoting angiogenesis gene regulation, which may promote the formation of blood vessels at hypoxia portion, and thus contribute to bone formation. <br> OBJECTIVE:To construct the Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) lentiviral eukaryotic expression vectors and to detect their impact on the osteogenic gene expression of rabbit bone marrow mesenchymal stem cells. <br> METHODS:The Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) lentiviral eukaryotic expression vectors were constructed according to the wild type human HIF-1αgene sequence and the determined restriction sites of human HIF-1αpoint mutant sequence. Rabbit bone marrow mesenchymal stem cells were transfected with the prepared Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) virus solution. <br> RESULTS AND CONCLUSION:Immunofluorescence microscopy observations indicated that the cells of Lenti-HIF-1α-IRES-EGFP (wild type) group and Lenti-HIF-1α-IRES-EGFP (point mutant type) group showed no obvious fluorescence on transfected 7 days, and two groups of cells showed a more obvious green fluorescent after transfection 14 days. Quantitative PCR analysis results showed that there were obvious HIF-1αand bonemorphogenetic protein 2 gene expressions on days 7 after transfection and the two genes stil showed highly expression levels after transfection 14 days. The two lentiviral eukaryotic expression vectors of Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) could be constructed according to the wild type human HIF-1αgene sequence and the determined restriction sites of human HIF-1αpoint mutant sequence;HIF-1αgene can promote the osteogenic gene expression of lentivirus-transfected rabbit bone marrow mesenchymal stem cells.关键词
干细胞/骨髓干细胞/低氧诱导因子1α/点突变/慢病毒/转染/骨髓间充质干细胞/基因治疗/聚合酶链反应/细胞学实验/骨形态发生蛋白2/国家自然科学基金Key words
lentivirus/mesenchymal stem cells/bone marrow/bone morphogenetic proteins/viruses/transfection分类
医药卫生引用本文复制引用
张劲娥,王淑红,张忻,郭华艳,郭娜,黄远亮..低氧诱导因子1α慢病毒载体转染骨髓间充质干细胞的成骨基因表达[J].中国组织工程研究,2014,(1):57-62,6.基金项目
国家自然科学基金重点项目(81070806)；浙江省自然科学基金(LQ12H14002)；上海市科委医学重点课题(10JC1413300)；杭州市医药卫生科技计划项目(2012A015)@@@@the National Natural Science Foundation of China, No.81070806 （81070806）
the Natural Science Foundation of Zhejiang Province, No. LQ12H14002 （）
Medical Key Topics of Shanghai Science and Technology Commission, No.10JC1413300 （）
the Medicine and Health Science and Technology Projects in Hangzhou, No.2012A015 （）
