Microvascular research · 2016
Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells.
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Markiewicz Margaret, Panneerselvam Kavin, Marks Natalia
Аннотация
PURPOSE: To examine the possible role of Klotho (Kl) in human microvasculature.
METHODS: The expression level of Kl in primary human dermal microvascular endothelial cells (HDMECs) and primary human dermal fibroblasts (HFb) was detected by real-time polymerase chain reaction amplification (qRT-PCR), Western blot analyses and immunohistochemistry. Migration of HDMECs and HFb was examined in monolayer wound healing "scratch assay" and Transwell assay. Proliferation of these cells was examined using Cell Proliferation BrdU incorporation assay.
RESULTS: Our results have shown that downregulation of Kl abrogated HDMECs migration after 48h. On the other hand, migration of HFb significantly increased after blocking Kl. Lack of Kl decreased expression of genes involved in the activation of endothelial cells and enhanced expression of genes involved in extracellular matrix remodeling and organization of connective tissue.
CONCLUSIONS: This study for the first time provides the evidence that Kl is expressed in HDMECs and HFb. Additionally, we have demonstrated that Kl is implicated in the process of angiogenesis of human dermal microvasculature.
METHODS: The expression level of Kl in primary human dermal microvascular endothelial cells (HDMECs) and primary human dermal fibroblasts (HFb) was detected by real-time polymerase chain reaction amplification (qRT-PCR), Western blot analyses and immunohistochemistry. Migration of HDMECs and HFb was examined in monolayer wound healing "scratch assay" and Transwell assay. Proliferation of these cells was examined using Cell Proliferation BrdU incorporation assay.
RESULTS: Our results have shown that downregulation of Kl abrogated HDMECs migration after 48h. On the other hand, migration of HFb significantly increased after blocking Kl. Lack of Kl decreased expression of genes involved in the activation of endothelial cells and enhanced expression of genes involved in extracellular matrix remodeling and organization of connective tissue.
CONCLUSIONS: This study for the first time provides the evidence that Kl is expressed in HDMECs and HFb. Additionally, we have demonstrated that Kl is implicated in the process of angiogenesis of human dermal microvasculature.
Предметные рубрики MeSH
Cell MovementCell ProliferationCells, CulturedEndothelial CellsFibroblastsForeskinGene Expression ProfilingGene Expression RegulationGlucuronidaseHumansInfant, NewbornKlotho ProteinsMaleMicrovesselsNeovascularization, PhysiologicOligonucleotide Array Sequence AnalysisRNA InterferenceSignal TransductionSkinTime FactorsTransfection