HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo

Mijeong Jeon, Yooseok Shin, Jaeeun Jung, Ui Won Jung, Jae Hoon Lee, Jae Seung Moon, Ilkoo Kim, Jin Su Shin, Sang Kyou Lee, Je Seon Song

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)99-107
Number of pages9
JournalMolecular and Cellular Biochemistry
Volume437
Issue number1-2
DOIs
Publication statusPublished - 2018 Jan 1

Fingerprint

Hypoxia-Inducible Factor 1
DNA-Binding Proteins
Umbilical Veins
Vascular Endothelium
Genes
Polymerase chain reaction
In Vitro Techniques
Real-Time Polymerase Chain Reaction
Assays
Hemoglobins
Staining and Labeling
Gene Expression
Proteins
Blood vessels
Hematoxylin
Eosine Yellowish-(YS)
Gene expression
Vascular Endothelial Growth Factor A
Blood Vessels

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

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. / HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo. In: Molecular and Cellular Biochemistry. 2018 ; Vol. 437, No. 1-2. pp. 99-107.
@article{202e996a421c412180a8b8500cbcf6c6,
title = "HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo",
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.",
author = "Mijeong Jeon and Yooseok Shin and Jaeeun Jung and Jung, {Ui Won} and Lee, {Jae Hoon} and Moon, {Jae Seung} and Ilkoo Kim and Shin, {Jin Su} and Lee, {Sang Kyou} and Song, {Je Seon}",
year = "2018",
month = "1",
day = "1",
doi = "10.1007/s11010-017-3098-6",
language = "English",
volume = "437",
pages = "99--107",
journal = "Molecular and Cellular Biochemistry",
issn = "0300-8177",
publisher = "Springer Netherlands",
number = "1-2",

}

HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo. / 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.

In: Molecular and Cellular Biochemistry, Vol. 437, No. 1-2, 01.01.2018, p. 99-107.

Research output: Contribution to journalArticle

TY - JOUR

T1 - HIF1A overexpression using cell-penetrating DNA-binding protein induces angiogenesis in vitro and in vivo

AU - Jeon, Mijeong

AU - Shin, Yooseok

AU - Jung, Jaeeun

AU - Jung, Ui Won

AU - Lee, Jae Hoon

AU - Moon, Jae Seung

AU - Kim, Ilkoo

AU - Shin, Jin Su

AU - Lee, Sang Kyou

AU - Song, Je Seon

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85025083208&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85025083208&partnerID=8YFLogxK

U2 - 10.1007/s11010-017-3098-6

DO - 10.1007/s11010-017-3098-6

M3 - Article

C2 - 28660411

AN - SCOPUS:85025083208

VL - 437

SP - 99

EP - 107

JO - Molecular and Cellular Biochemistry

JF - Molecular and Cellular Biochemistry

SN - 0300-8177

IS - 1-2

ER -