HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo
Hypoxia-inducible factor-1 alpha ( HIF1A ) is an important transcription factor for angiogenesis. Recent studies have used the protein transduction domain (PTD) to deliver genes, but the PTD has not been used to induce the expression of HIF1A . This study aimed at using a novel PTD (Hph-1-GAL4; ARVR...
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Veröffentlicht in: | Molecular and cellular biochemistry 2018, Vol.437 (1-2), p.99-107 |
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container_title | Molecular and cellular biochemistry |
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creator | Jeon, Mijeong Shin, Yooseok Jung, Jaeeun Jung, Ui-Won Lee, Jae-Hoon Moon, Jae-Seung Kim, Ilkoo Shin, Jin-Su Lee, Sang-Kyou Song, Je Seon |
description | Hypoxia-inducible factor-1 alpha (
HIF1A
) is an important transcription factor for angiogenesis. Recent studies have used the protein transduction domain (PTD) to deliver genes, but the PTD has not been used to induce the expression of
HIF1A
. This study aimed at using a novel PTD (Hph-1-GAL4; ARVRRRGPRR) to overexpress the
HIF1A
and identify the effects on angiogenesis in vitro and in vivo. Overexpression of
HIF1A
was induced using Hph-1-GAL4 in human umbilical vein/vascular endothelium cells (HUVEC). The expression levels of genes were analyzed by the quantitative real-time polymerase chain reaction (qPCR) after 2 and 4 days, respectively. An in vitro tube formation was performed using Diff-Quik staining.
HIF1A
and Hph-1-GAL4 were injected subcutaneously into the ventral area of each 5-week-old mouse. All of the plugs were retrieved after 1 week, and the gene expression levels were evaluated by qPCR. Each Matrigel plug was evaluated using the hemoglobin assay and hematoxylin and eosin (HE) staining. The expression levels of
HIF1A
and
HIF1A
target genes were significantly higher in
HIF1A
-transfected HUVEC than in control HUVEC in vitro. In the in vivo Matrigel plug assay, the amount of hemoglobin was significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. Blood vessels were identified in the
HIF1A
-treatment group. The expression levels of
HIF1A
, vascular endothelial growth factor (
Vegf)
, and
Cd31
were significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. These findings suggest that using Hph-1-G4D to overexpress
HIF1A
might be useful for transferring genes and regenerating tissues. |
doi_str_mv | 10.1007/s11010-017-3098-6 |
format | Article |
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HIF1A
) is an important transcription factor for angiogenesis. Recent studies have used the protein transduction domain (PTD) to deliver genes, but the PTD has not been used to induce the expression of
HIF1A
. This study aimed at using a novel PTD (Hph-1-GAL4; ARVRRRGPRR) to overexpress the
HIF1A
and identify the effects on angiogenesis in vitro and in vivo. Overexpression of
HIF1A
was induced using Hph-1-GAL4 in human umbilical vein/vascular endothelium cells (HUVEC). The expression levels of genes were analyzed by the quantitative real-time polymerase chain reaction (qPCR) after 2 and 4 days, respectively. An in vitro tube formation was performed using Diff-Quik staining.
HIF1A
and Hph-1-GAL4 were injected subcutaneously into the ventral area of each 5-week-old mouse. All of the plugs were retrieved after 1 week, and the gene expression levels were evaluated by qPCR. Each Matrigel plug was evaluated using the hemoglobin assay and hematoxylin and eosin (HE) staining. The expression levels of
HIF1A
and
HIF1A
target genes were significantly higher in
HIF1A
-transfected HUVEC than in control HUVEC in vitro. In the in vivo Matrigel plug assay, the amount of hemoglobin was significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. Blood vessels were identified in the
HIF1A
-treatment group. The expression levels of
HIF1A
, vascular endothelial growth factor (
Vegf)
, and
Cd31
were significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. These findings suggest that using Hph-1-G4D to overexpress
HIF1A
might be useful for transferring genes and regenerating tissues.</description><identifier>ISSN: 0300-8177</identifier><identifier>EISSN: 1573-4919</identifier><identifier>DOI: 10.1007/s11010-017-3098-6</identifier><identifier>PMID: 28660411</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Analysis ; Angiogenesis ; Biochemistry ; Biomedical and Life Sciences ; Blood vessels ; Cardiology ; Cells ; Deoxyribonucleic acid ; DNA ; DNA-binding protein ; Endothelium ; Gene expression ; Genes ; Hemoglobin ; Hemoglobins ; Hypoxia ; Hypoxia-inducible factor 1 ; Hypoxia-inducible factor 1a ; Life Sciences ; Medical Biochemistry ; Oncology ; Plugs ; Polymerase chain reaction ; Protein binding ; Proteins ; Staining ; Umbilical vein ; Vascular endothelial growth factor</subject><ispartof>Molecular and cellular biochemistry, 2018, Vol.437 (1-2), p.99-107</ispartof><rights>Springer Science+Business Media, LLC 2017</rights><rights>COPYRIGHT 2018 Springer</rights><rights>Molecular and Cellular Biochemistry is a copyright of Springer, (2017). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c439t-dd002ce3f233f5e1c4e25e4cd7babe005d44726a7935d9b92af09ee77b150e283</citedby><cites>FETCH-LOGICAL-c439t-dd002ce3f233f5e1c4e25e4cd7babe005d44726a7935d9b92af09ee77b150e283</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11010-017-3098-6$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11010-017-3098-6$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28660411$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jeon, Mijeong</creatorcontrib><creatorcontrib>Shin, Yooseok</creatorcontrib><creatorcontrib>Jung, Jaeeun</creatorcontrib><creatorcontrib>Jung, Ui-Won</creatorcontrib><creatorcontrib>Lee, Jae-Hoon</creatorcontrib><creatorcontrib>Moon, Jae-Seung</creatorcontrib><creatorcontrib>Kim, Ilkoo</creatorcontrib><creatorcontrib>Shin, Jin-Su</creatorcontrib><creatorcontrib>Lee, Sang-Kyou</creatorcontrib><creatorcontrib>Song, Je Seon</creatorcontrib><title>HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo</title><title>Molecular and cellular biochemistry</title><addtitle>Mol Cell Biochem</addtitle><addtitle>Mol Cell Biochem</addtitle><description>Hypoxia-inducible factor-1 alpha (
HIF1A
) is an important transcription factor for angiogenesis. Recent studies have used the protein transduction domain (PTD) to deliver genes, but the PTD has not been used to induce the expression of
HIF1A
. This study aimed at using a novel PTD (Hph-1-GAL4; ARVRRRGPRR) to overexpress the
HIF1A
and identify the effects on angiogenesis in vitro and in vivo. Overexpression of
HIF1A
was induced using Hph-1-GAL4 in human umbilical vein/vascular endothelium cells (HUVEC). The expression levels of genes were analyzed by the quantitative real-time polymerase chain reaction (qPCR) after 2 and 4 days, respectively. An in vitro tube formation was performed using Diff-Quik staining.
HIF1A
and Hph-1-GAL4 were injected subcutaneously into the ventral area of each 5-week-old mouse. All of the plugs were retrieved after 1 week, and the gene expression levels were evaluated by qPCR. Each Matrigel plug was evaluated using the hemoglobin assay and hematoxylin and eosin (HE) staining. The expression levels of
HIF1A
and
HIF1A
target genes were significantly higher in
HIF1A
-transfected HUVEC than in control HUVEC in vitro. In the in vivo Matrigel plug assay, the amount of hemoglobin was significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. Blood vessels were identified in the
HIF1A
-treatment group. The expression levels of
HIF1A
, vascular endothelial growth factor (
Vegf)
, and
Cd31
were significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. These findings suggest that using Hph-1-G4D to overexpress
HIF1A
might be useful for transferring genes and regenerating tissues.</description><subject>Analysis</subject><subject>Angiogenesis</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Blood vessels</subject><subject>Cardiology</subject><subject>Cells</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA-binding protein</subject><subject>Endothelium</subject><subject>Gene expression</subject><subject>Genes</subject><subject>Hemoglobin</subject><subject>Hemoglobins</subject><subject>Hypoxia</subject><subject>Hypoxia-inducible factor 1</subject><subject>Hypoxia-inducible factor 1a</subject><subject>Life Sciences</subject><subject>Medical Biochemistry</subject><subject>Oncology</subject><subject>Plugs</subject><subject>Polymerase chain reaction</subject><subject>Protein binding</subject><subject>Proteins</subject><subject>Staining</subject><subject>Umbilical vein</subject><subject>Vascular endothelial growth factor</subject><issn>0300-8177</issn><issn>1573-4919</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1kU1v1DAQhi0EosvCD-CCInHpxWXGduL4uCqUVqrgAmcrH5PI1a692Mmq_HscpeVLIB_sef3Mq7Ffxl4jXCCAfpcQAYEDai7B1Lx6wjZYasmVQfOUbUAC8Bq1PmMvUrqDDAPic3Ym6qoChbhh7vrmCndFOFGk-2OklFzwxZycH4uO9nt-JE9TbKZFeP9px1vn--V8jGEi54tczh2lovGjC2OGk0tZLE5uiiGr_Vqcwkv2bGj2iV497Fv29erDl8trfvv5483l7pZ3SpqJ9z2A6EgOQsqhJOwUiZJU1-u2aQmg7JXSomq0kWVvWiOaAQyR1i2WQKKWW3a--uYJv82UJntwaXlK4ynMyaJBVStjssGWvf0LvQtz9Hm6TNUKZF0i_qLGZk_W-SHk_-gWU7srBcqyMkJl6uIfVF49HVwXPA0u63804NrQxZBSpMEeozs08btFsEu8do3X5njtEq-tcs-bh4Hn9kD9z47HPDMgViDlKz9S_O1F_3X9AW2jrig</recordid><startdate>2018</startdate><enddate>2018</enddate><creator>Jeon, Mijeong</creator><creator>Shin, 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B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QP</scope><scope>7T5</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>2018</creationdate><title>HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo</title><author>Jeon, Mijeong ; Shin, Yooseok ; Jung, Jaeeun ; Jung, Ui-Won ; Lee, Jae-Hoon ; Moon, Jae-Seung ; Kim, Ilkoo ; Shin, Jin-Su ; Lee, Sang-Kyou ; Song, Je Seon</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c439t-dd002ce3f233f5e1c4e25e4cd7babe005d44726a7935d9b92af09ee77b150e283</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Analysis</topic><topic>Angiogenesis</topic><topic>Biochemistry</topic><topic>Biomedical and Life Sciences</topic><topic>Blood vessels</topic><topic>Cardiology</topic><topic>Cells</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA-binding protein</topic><topic>Endothelium</topic><topic>Gene expression</topic><topic>Genes</topic><topic>Hemoglobin</topic><topic>Hemoglobins</topic><topic>Hypoxia</topic><topic>Hypoxia-inducible factor 1</topic><topic>Hypoxia-inducible factor 1a</topic><topic>Life Sciences</topic><topic>Medical Biochemistry</topic><topic>Oncology</topic><topic>Plugs</topic><topic>Polymerase chain reaction</topic><topic>Protein binding</topic><topic>Proteins</topic><topic>Staining</topic><topic>Umbilical vein</topic><topic>Vascular endothelial growth factor</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jeon, Mijeong</creatorcontrib><creatorcontrib>Shin, Yooseok</creatorcontrib><creatorcontrib>Jung, Jaeeun</creatorcontrib><creatorcontrib>Jung, Ui-Won</creatorcontrib><creatorcontrib>Lee, Jae-Hoon</creatorcontrib><creatorcontrib>Moon, Jae-Seung</creatorcontrib><creatorcontrib>Kim, Ilkoo</creatorcontrib><creatorcontrib>Shin, Jin-Su</creatorcontrib><creatorcontrib>Lee, Sang-Kyou</creatorcontrib><creatorcontrib>Song, Je 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Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular and cellular biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jeon, Mijeong</au><au>Shin, Yooseok</au><au>Jung, Jaeeun</au><au>Jung, Ui-Won</au><au>Lee, Jae-Hoon</au><au>Moon, Jae-Seung</au><au>Kim, Ilkoo</au><au>Shin, Jin-Su</au><au>Lee, Sang-Kyou</au><au>Song, Je Seon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo</atitle><jtitle>Molecular and cellular biochemistry</jtitle><stitle>Mol Cell Biochem</stitle><addtitle>Mol Cell Biochem</addtitle><date>2018</date><risdate>2018</risdate><volume>437</volume><issue>1-2</issue><spage>99</spage><epage>107</epage><pages>99-107</pages><issn>0300-8177</issn><eissn>1573-4919</eissn><abstract>Hypoxia-inducible factor-1 alpha (
HIF1A
) is an important transcription factor for angiogenesis. Recent studies have used the protein transduction domain (PTD) to deliver genes, but the PTD has not been used to induce the expression of
HIF1A
. This study aimed at using a novel PTD (Hph-1-GAL4; ARVRRRGPRR) to overexpress the
HIF1A
and identify the effects on angiogenesis in vitro and in vivo. Overexpression of
HIF1A
was induced using Hph-1-GAL4 in human umbilical vein/vascular endothelium cells (HUVEC). The expression levels of genes were analyzed by the quantitative real-time polymerase chain reaction (qPCR) after 2 and 4 days, respectively. An in vitro tube formation was performed using Diff-Quik staining.
HIF1A
and Hph-1-GAL4 were injected subcutaneously into the ventral area of each 5-week-old mouse. All of the plugs were retrieved after 1 week, and the gene expression levels were evaluated by qPCR. Each Matrigel plug was evaluated using the hemoglobin assay and hematoxylin and eosin (HE) staining. The expression levels of
HIF1A
and
HIF1A
target genes were significantly higher in
HIF1A
-transfected HUVEC than in control HUVEC in vitro. In the in vivo Matrigel plug assay, the amount of hemoglobin was significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. Blood vessels were identified in the
HIF1A
-treatment group. The expression levels of
HIF1A
, vascular endothelial growth factor (
Vegf)
, and
Cd31
were significantly higher in the
HIF1A
-treatment group than in the PBS-treatment group. These findings suggest that using Hph-1-G4D to overexpress
HIF1A
might be useful for transferring genes and regenerating tissues.</abstract><cop>New York</cop><pub>Springer US</pub><pmid>28660411</pmid><doi>10.1007/s11010-017-3098-6</doi><tpages>9</tpages></addata></record> |
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ispartof | Molecular and cellular biochemistry, 2018, Vol.437 (1-2), p.99-107 |
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source | SpringerLink Journals (MCLS) |
subjects | Analysis Angiogenesis Biochemistry Biomedical and Life Sciences Blood vessels Cardiology Cells Deoxyribonucleic acid DNA DNA-binding protein Endothelium Gene expression Genes Hemoglobin Hemoglobins Hypoxia Hypoxia-inducible factor 1 Hypoxia-inducible factor 1a Life Sciences Medical Biochemistry Oncology Plugs Polymerase chain reaction Protein binding Proteins Staining Umbilical vein Vascular endothelial growth factor |
title | HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo |
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