Mutant hypoxia inducible factor 1alpha transfection promotes proliferation of bone marrow mesenchymal stem cells

Zhang Wei-wei; Wang Jue

doi:10.3969/j.issn.2095-4344.2017.25.004

Abstract：BACKGROUND: It has been found that hypoxia inducible 1α (HIF1α) gene can improve the ability of tissues and cells to survive in the ischemic environment, which is of great significance in promoting blood vessel regeneration.OBJECTIVE: To investigate the effect of the recombinant adenovirus Ad-CMV-HIF1αmu-IRES-hrGFP-1 on the proliferation of bone marrow mesenchymal stem cells.METHODS: Passage 3 bone marrow mesenchymal stem cells were cultured and transfected with Ad-CMV-HIF1αmu-IRES-hrGFP-1 virus (experimental group), with non-mutated gene Ad-CMV-HIF1α-IRES-hrGFP (positive control group), with Ad-CMV-IRES-hrGFP-1 empty virus (negative control group), or with no virus solution (blank control group). After transfection for 24 hours, the expression of HIF1α mRNA and protein was detected. Cell proliferation was detected by MTT assay at 24, 48, 72, 96 and 120 hours after transfection.RESULTS AND CONCLUSION: (1) Except that the mRNA expression of HIF1α in the experimental group and the positive control group was significantly higher than that in the negative control group and the blank control group (P < 0.05), there was no significant difference between two groups. (2) The expression of HIF1α protein in the experimental group was significantly higher than that in the positive control group, the negative control group and the blank control group (P < 0.05), and there was no significant difference between the latter three groups. (3) Cell proliferation was faster in the experimental group than the positive control group and the blank control group (P < 0.05), and moreover, there was no significant difference between the latter two groups. Our experimental findings indicate that the recombinant adenovirus-mediated three-point mutant HIF1α combined with hrGFP gene transfected bone marrow mesenchymal stem cells could not only express the target protein, HIF1α, under normoxic conditions, but also promote the proliferation of bone marrow mesenchymal stem cells.