The effects of marrow mesenchymal stem cells transfected with hypoxia-inducible factor-1α gene

Objective To construct hypoxia-inducible factor 1 αteukaryotic expression vector-with pcDNA3.1 and investigate its transfective efficiency into mesenchymal stem cells (MSCs) in vitro and the expression of HIF-1α gene in MSCs. Methods mRNA of Wistar Rats'myocardial ceils was extracted, and cDNA was synthesized with reverse transcription kit. HIF-1α was amplified by polymerase chain reaction (PCR), and constructed into pcDNA3.1. HIF-1α-pcDNA3.1 was transfected into MSCs by liposome mediated method. The expression of HIF-1 α in the cells was detected by Western Blot Analysis and ELISA. Results Eukaryotic expression vector HIF-1α-pcDNA3.1 was constructed successfully. In MSCs transfected with HIF-1α gene , both the expression of HIF-1α mRNA and HIF-1α protein were obviously increased. Conclusion HIF-1α was successfully cloned. HIF-1α-pcDNA3.1 can be effectively transfected into MSCs with liposome-mediated method, which can result stable expression of HIF-1αin transfected MSCs. Key words: DNA-binding proteins/GE ;  Cloning;  molecular;  Myeloid progenitor cells