Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells.
To examine the possible role of Klotho (Kl) in human microvasculature.



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.



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.



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.
Division of Rheumatology and Immunology, Medical University of South Carolina, Charleston, SC 29425, United States. Electronic address: markiewi@musc.edu.
Margaret
M
Markiewicz
Division of Rheumatology and Immunology, Medical University of South Carolina, Charleston, SC 29425, United States.
Kavin
K
Panneerselvam
NYP - Columbia University Medical Center, New York, NY 10032, United States.
Natalia
N
Marks
D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D052816:Foreskin / Q000098:blood supply*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007231:Infant, Newborn; D000090265:Klotho Proteins; D008297:Male; D055806:Microvessels / Q000166:cytology*; D018919:Neovascularization, Physiologic; D020411:Oligonucleotide Array Sequence Analysis; D034622:RNA Interference; D015398:Signal Transduction; D012867:Skin / Q000098:blood supply*; D013997:Time Factors; D014162:Transfection
D005966:Glucuronidase; D000090265:Klotho Proteins
Angiogenesis; Dermal fibroblasts; Dermal microvascular endothelial cells; Klotho