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 anal...
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| Veröffentlicht in: | Microvascular research 2016-09, Vol.107, p.76-82 |
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| creator | Markiewicz, Margaret Panneerselvam, Kavin Marks, Natalia |
| description | 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.
•Klotho is expressed in microvascular endothelial cells and dermal fibroblasts.•Lack of Kl abrogates migration of human dermal microvascular endothelial cells.•Lack of Kl enhances migration of human dermal fibroblasts.•Decrease in gene expression involved in the activation of endothelial cells. |
| doi_str_mv | 10.1016/j.mvr.2016.05.005 |
| format | Article |
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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.
•Klotho is expressed in microvascular endothelial cells and dermal fibroblasts.•Lack of Kl abrogates migration of human dermal microvascular endothelial cells.•Lack of Kl enhances migration of human dermal fibroblasts.•Decrease in gene expression involved in the activation of endothelial cells.</description><identifier>ISSN: 0026-2862</identifier><identifier>EISSN: 1095-9319</identifier><identifier>DOI: 10.1016/j.mvr.2016.05.005</identifier><identifier>PMID: 27260080</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Angiogenesis ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Dermal fibroblasts ; Dermal microvascular endothelial cells ; Endothelial Cells - metabolism ; Fibroblasts - metabolism ; Foreskin - blood supply ; Gene Expression Profiling - methods ; Gene Expression Regulation ; Glucuronidase - genetics ; Glucuronidase - metabolism ; Humans ; Infant, Newborn ; Klotho ; Male ; Microvessels - cytology ; Neovascularization, Physiologic ; Oligonucleotide Array Sequence Analysis ; RNA Interference ; Signal Transduction ; Skin - blood supply ; Time Factors ; Transfection</subject><ispartof>Microvascular research, 2016-09, Vol.107, p.76-82</ispartof><rights>2016</rights><rights>Published by Elsevier Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c517t-110f11c3cfe825c427bec5adb60bf09e59c0fb72d12a5710ea0741e5e5e0de103</citedby><cites>FETCH-LOGICAL-c517t-110f11c3cfe825c427bec5adb60bf09e59c0fb72d12a5710ea0741e5e5e0de103</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.mvr.2016.05.005$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,777,781,882,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27260080$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Markiewicz, Margaret</creatorcontrib><creatorcontrib>Panneerselvam, Kavin</creatorcontrib><creatorcontrib>Marks, Natalia</creatorcontrib><title>Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells</title><title>Microvascular research</title><addtitle>Microvasc Res</addtitle><description>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.
•Klotho is expressed in microvascular endothelial cells and dermal fibroblasts.•Lack of Kl abrogates migration of human dermal microvascular endothelial cells.•Lack of Kl enhances migration of human dermal fibroblasts.•Decrease in gene expression involved in the activation of endothelial cells.</description><subject>Angiogenesis</subject><subject>Cell Movement</subject><subject>Cell Proliferation</subject><subject>Cells, Cultured</subject><subject>Dermal fibroblasts</subject><subject>Dermal microvascular endothelial cells</subject><subject>Endothelial Cells - metabolism</subject><subject>Fibroblasts - metabolism</subject><subject>Foreskin - blood supply</subject><subject>Gene Expression Profiling - methods</subject><subject>Gene Expression Regulation</subject><subject>Glucuronidase - genetics</subject><subject>Glucuronidase - metabolism</subject><subject>Humans</subject><subject>Infant, Newborn</subject><subject>Klotho</subject><subject>Male</subject><subject>Microvessels - cytology</subject><subject>Neovascularization, Physiologic</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>RNA Interference</subject><subject>Signal Transduction</subject><subject>Skin - blood supply</subject><subject>Time Factors</subject><subject>Transfection</subject><issn>0026-2862</issn><issn>1095-9319</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1r3DAQhkVJabZpf0Auwcdc7M7IK38QKJTQLxoolPZYhCyNs1pkaSPZC_331bKbkF6KDiM0z7yjmZexS4QKAZt322rax4rnawWiAhAv2AqhF2VfY3_GVgC8KXnX8HP2OqUtAKLo-St2zlveAHSwYr9_BEdFGItvLsybUFhfTPY-qtkGXyhvil0Mzo50esngZpmULwzFSbnM6hj2KunFqViQN1mEnM0ZTc6lN-zlqFyit6d4wX59-vjz9kt59_3z19sPd6UW2M4lIoyIutYjdVzoNW8H0kKZoYFhhJ5Er2EcWm6QK9EikIJ2jSTyAUMI9QV7f9TdLcNERpOfo3JyF-2k4h8ZlJX_ZrzdyPuwl-u-7tr1QeD6JBDDw0JplpNNhxGUp7AkiR3wtu6QdxnFI5onTynS-NQGQR5skVuZbZEHWyQImW3JNVfP__dU8ehDBm6OAOUt7S1FmbQlr8nYSHqWJtj_yP8Fhkyghw</recordid><startdate>20160901</startdate><enddate>20160901</enddate><creator>Markiewicz, Margaret</creator><creator>Panneerselvam, Kavin</creator><creator>Marks, Natalia</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20160901</creationdate><title>Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells</title><author>Markiewicz, Margaret ; Panneerselvam, Kavin ; Marks, Natalia</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c517t-110f11c3cfe825c427bec5adb60bf09e59c0fb72d12a5710ea0741e5e5e0de103</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Angiogenesis</topic><topic>Cell Movement</topic><topic>Cell Proliferation</topic><topic>Cells, Cultured</topic><topic>Dermal fibroblasts</topic><topic>Dermal microvascular endothelial cells</topic><topic>Endothelial Cells - metabolism</topic><topic>Fibroblasts - metabolism</topic><topic>Foreskin - blood supply</topic><topic>Gene Expression Profiling - methods</topic><topic>Gene Expression Regulation</topic><topic>Glucuronidase - genetics</topic><topic>Glucuronidase - metabolism</topic><topic>Humans</topic><topic>Infant, Newborn</topic><topic>Klotho</topic><topic>Male</topic><topic>Microvessels - cytology</topic><topic>Neovascularization, Physiologic</topic><topic>Oligonucleotide Array Sequence Analysis</topic><topic>RNA Interference</topic><topic>Signal Transduction</topic><topic>Skin - blood supply</topic><topic>Time Factors</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Markiewicz, Margaret</creatorcontrib><creatorcontrib>Panneerselvam, Kavin</creatorcontrib><creatorcontrib>Marks, Natalia</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Microvascular research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Markiewicz, Margaret</au><au>Panneerselvam, Kavin</au><au>Marks, Natalia</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells</atitle><jtitle>Microvascular research</jtitle><addtitle>Microvasc Res</addtitle><date>2016-09-01</date><risdate>2016</risdate><volume>107</volume><spage>76</spage><epage>82</epage><pages>76-82</pages><issn>0026-2862</issn><eissn>1095-9319</eissn><abstract>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.
•Klotho is expressed in microvascular endothelial cells and dermal fibroblasts.•Lack of Kl abrogates migration of human dermal microvascular endothelial cells.•Lack of Kl enhances migration of human dermal fibroblasts.•Decrease in gene expression involved in the activation of endothelial cells.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>27260080</pmid><doi>10.1016/j.mvr.2016.05.005</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Angiogenesis Cell Movement Cell Proliferation Cells, Cultured Dermal fibroblasts Dermal microvascular endothelial cells Endothelial Cells - metabolism Fibroblasts - metabolism Foreskin - blood supply Gene Expression Profiling - methods Gene Expression Regulation Glucuronidase - genetics Glucuronidase - metabolism Humans Infant, Newborn Klotho Male Microvessels - cytology Neovascularization, Physiologic Oligonucleotide Array Sequence Analysis RNA Interference Signal Transduction Skin - blood supply Time Factors Transfection |
| title | Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells |
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