Genetically modified endothelial progenitor cells with hNotch1.ICN overexpression display facilitated angiogenesis.

Abstract

BackgroundThis study focused on hNotch1.ICN overexpression and investigated how it affects the biological behavior of endothelial progenitor cells (EPC) in vitro.MethodsCCK 8 assay was used to evaluate overexpressed hNotch1.ICN to determine how to influence EPCs’ survivability. The Annexin V/PI method was used to detect overexpressed hNotch1.ICN and to influence EPC apoptosis. A flow cytometry instrument was used to assess the overexpression of hNotch1.ICN and determine how to influence the EPC cell cycle. Transwell was used to investigate how overexpressed hNotch1.ICN EPCs migrate using their endothelial ability and adhesive ability with activated endothelial cells and angiogenesis ability. After lentivirus gene transfection, qPCR and Western blot were used to detect a notch signaling pathway downstream of the signaling molecules Hes 1 and Hey 1 mRNA and protein expression. The role of the Notch.1 intracellular domain as a candidate EPC regulator with its differential expression and Hes 1 and Hey 1 expression of Notch downstream signaling molecules in separate groups was analyzed.ResultsA detailed analysis revealed an over-expressed hNotch1.ICN gene had no significant effect on canine EPC growth, strengthened EPC antiapoptotic ability, increased numbers of EPCs that underwent cell cycle arrest in the G2 phase, inhibited EPCs differentiation, and enhanced Hes 1 and Hey 1 expression. Moreover, an over-expressed hNotch1 ICN gene promotes EPCs to migrate across ECs, promotes EPCs to adhere to activating endothelial cells, and induces angiogenesis in vitro.ConclusionsOver-expressed hNotch1.ICN onto EPCs could be used as a potential candidate to treat many ischemic diseases.