Influence of hypoxia-inducible factor-1 alpha lentiviral vector on osteogenic gene expression of rabbit bone marrow mesenchymal stem cells / 中国组织工程研究
Chinese Journal of Tissue Engineering Research
;
(53): 57-62, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-443648
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.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.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. RESULTS ANDCONCLUSION:
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Tissue Engineering Research
Year:
2014
Type:
Article
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