1. Transfection of hypoxia-inducible factor-1α mRNA upregulates the expression of genes encoding angiogenic growth factors.
- Author
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Wlodarczyk J, Leng A, Abadchi SN, Shababi N, Mokhtari-Esbuie F, Gheshlaghi S, Ravari MR, Pippenger EK, Afrasiabi A, Ha J, Abraham JM, and Harmon JW
- Subjects
- Humans, RNA, Messenger metabolism, Transfection, Intercellular Signaling Peptides and Proteins genetics, Protein Isoforms genetics, Vascular Endothelial Growth Factor A metabolism, Hypoxia-Inducible Factor 1, alpha Subunit genetics
- Abstract
Hypoxia-Inducible Factor-1α (HIF-1α) has presented a new direction for ischemic preconditioning of surgical flaps to promote their survival. In a previous study, we demonstrated the effectiveness of HIF-1a DNA plasmids in this application. In this study, to avoid complications associated with plasmid use, we sought to express HIF-1α through mRNA transfection and determine its biological activity by measuring the upregulation of downstream angiogenic genes. We transfected six different HIF-1a mRNAs-one predominant, three variant, and two novel mutant isoforms-into primary human dermal fibroblasts using Lipofectamine, and assessed mRNA levels using RT-qPCR. At all time points examined after transfection (3, 6, and 10 h), the levels of HIF-1α transcript were significantly higher in all HIF-1α transfected cells relative to the control (all p < 0.05, unpaired Student's T-test). Importantly, the expression of HIF-1α transcription response genes (VEGF, ANG-1, PGF, FLT1, and EDN1) was significantly higher in the cells transfected with all isoforms than with the control at six and/or ten hours post-transfection. All isoforms were transfected successfully into human fibroblast cells, resulting in the rapid upregulation of all five downstream angiogenic targets tested. These findings support the potential use of HIF-1α mRNA for protecting ischemic dermal flaps., (© 2024. The Author(s).)
- Published
- 2024
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