Downregulation of PTEN at corneal wound sites accelerates wound healing through increased cell migration

Lin Cao, Enrique O Graue-Hernandez, Vu Tran, Brian Reid, Jin Pu, Mark J Mannis, Min Zhao

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

PURPOSE: The PI3K/Akt pathway is required for cell polarization and migration, whereas the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) has inhibitory effects on the PI3K/Akt pathway. The authors therefore hypothesized that wounding would downregulate PTEN and that this downregulation would enhance wound healing.

METHODS: In human corneal epithelial (HCE) cell monolayer and rat cornea scratch wound models, the authors investigated PTEN and Akt expression using Western blot and immunofluorescence analyses. The effects of PTEN and PI3K inhibitors dipotassium bisperoxo (picolinato) oxovanadate (bpv(pic)) and LY294002 on cell migration and wound closure were investigated using time-lapse imaging. Finally, the authors investigated the effect of PTEN inhibition on wound healing in whole rat eyes.

RESULTS: In HCE cell monolayer and rat cornea, PTEN was downregulated at the wound edges within 30 minutes of wounding. The downregulation of PTEN was causal in a simultaneous increase in Akt activation, which was responsible for a significant increase in individual cell migration rate from 8.8 μm/h to 17.3 μm/h. An increased migration rate was maintained for 20 hours. PTEN inhibition significantly enhanced the wound healing rate in the HCE cell monolayer from 10 minutes onward after treatment and reduced the healing time in eye organ culture from 30 to 20 hours.

CONCLUSIONS: Injury to the corneal epithelium downregulates the expression of PTEN at wound edges, allowing increased PI3K/Akt signaling, thereby contributing to a significant enhancement of cell migration and wound healing. These results suggest that PTEN inhibition may be an effective treatment for corneal injury.

Original languageEnglish
Pages (from-to)2272-2278
Number of pages7
JournalInvestigative Ophthalmology & Visual Science
Volume52
Issue number5
DOIs
Publication statusPublished - Apr 2011

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Wound Healing
Cell Movement
Phosphatidylinositol 3-Kinases
Down-Regulation
Wounds and Injuries
Epithelial Cells
Cornea
Time-Lapse Imaging
Chromosomes, Human, Pair 10
Corneal Epithelium
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Organ Culture Techniques
Phosphoric Monoester Hydrolases
Fluorescent Antibody Technique
Western Blotting
Therapeutics

Keywords

  • animals
  • blotting, Western
  • cell movement
  • chromones
  • down-regulation
  • enzyme inhibitors
  • epithelium, corneal
  • female
  • fluorescent antibody technique, indirect
  • humans
  • male
  • morpholines
  • organ culture techniques
  • organometallic compounds
  • PTEN phosphohydrolase
  • phosphatidylinositol 3-kinases
  • proto-oncogene proteins c-akt
  • rats
  • rats, sprague-dawley
  • wound healing

Cite this

Downregulation of PTEN at corneal wound sites accelerates wound healing through increased cell migration. / Cao, Lin; Graue-Hernandez, Enrique O; Tran, Vu; Reid, Brian; Pu, Jin; Mannis, Mark J; Zhao, Min.

In: Investigative Ophthalmology & Visual Science, Vol. 52, No. 5, 04.2011, p. 2272-2278.

Research output: Contribution to journalArticle

Cao, Lin ; Graue-Hernandez, Enrique O ; Tran, Vu ; Reid, Brian ; Pu, Jin ; Mannis, Mark J ; Zhao, Min. / Downregulation of PTEN at corneal wound sites accelerates wound healing through increased cell migration. In: Investigative Ophthalmology & Visual Science. 2011 ; Vol. 52, No. 5. pp. 2272-2278.
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abstract = "PURPOSE: The PI3K/Akt pathway is required for cell polarization and migration, whereas the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) has inhibitory effects on the PI3K/Akt pathway. The authors therefore hypothesized that wounding would downregulate PTEN and that this downregulation would enhance wound healing.METHODS: In human corneal epithelial (HCE) cell monolayer and rat cornea scratch wound models, the authors investigated PTEN and Akt expression using Western blot and immunofluorescence analyses. The effects of PTEN and PI3K inhibitors dipotassium bisperoxo (picolinato) oxovanadate (bpv(pic)) and LY294002 on cell migration and wound closure were investigated using time-lapse imaging. Finally, the authors investigated the effect of PTEN inhibition on wound healing in whole rat eyes.RESULTS: In HCE cell monolayer and rat cornea, PTEN was downregulated at the wound edges within 30 minutes of wounding. The downregulation of PTEN was causal in a simultaneous increase in Akt activation, which was responsible for a significant increase in individual cell migration rate from 8.8 μm/h to 17.3 μm/h. An increased migration rate was maintained for 20 hours. PTEN inhibition significantly enhanced the wound healing rate in the HCE cell monolayer from 10 minutes onward after treatment and reduced the healing time in eye organ culture from 30 to 20 hours.CONCLUSIONS: Injury to the corneal epithelium downregulates the expression of PTEN at wound edges, allowing increased PI3K/Akt signaling, thereby contributing to a significant enhancement of cell migration and wound healing. These results suggest that PTEN inhibition may be an effective treatment for corneal injury.",
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TY - JOUR

T1 - Downregulation of PTEN at corneal wound sites accelerates wound healing through increased cell migration

AU - Cao, Lin

AU - Graue-Hernandez, Enrique O

AU - Tran, Vu

AU - Reid, Brian

AU - Pu, Jin

AU - Mannis, Mark J

AU - Zhao, Min

PY - 2011/4

Y1 - 2011/4

N2 - PURPOSE: The PI3K/Akt pathway is required for cell polarization and migration, whereas the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) has inhibitory effects on the PI3K/Akt pathway. The authors therefore hypothesized that wounding would downregulate PTEN and that this downregulation would enhance wound healing.METHODS: In human corneal epithelial (HCE) cell monolayer and rat cornea scratch wound models, the authors investigated PTEN and Akt expression using Western blot and immunofluorescence analyses. The effects of PTEN and PI3K inhibitors dipotassium bisperoxo (picolinato) oxovanadate (bpv(pic)) and LY294002 on cell migration and wound closure were investigated using time-lapse imaging. Finally, the authors investigated the effect of PTEN inhibition on wound healing in whole rat eyes.RESULTS: In HCE cell monolayer and rat cornea, PTEN was downregulated at the wound edges within 30 minutes of wounding. The downregulation of PTEN was causal in a simultaneous increase in Akt activation, which was responsible for a significant increase in individual cell migration rate from 8.8 μm/h to 17.3 μm/h. An increased migration rate was maintained for 20 hours. PTEN inhibition significantly enhanced the wound healing rate in the HCE cell monolayer from 10 minutes onward after treatment and reduced the healing time in eye organ culture from 30 to 20 hours.CONCLUSIONS: Injury to the corneal epithelium downregulates the expression of PTEN at wound edges, allowing increased PI3K/Akt signaling, thereby contributing to a significant enhancement of cell migration and wound healing. These results suggest that PTEN inhibition may be an effective treatment for corneal injury.

AB - PURPOSE: The PI3K/Akt pathway is required for cell polarization and migration, whereas the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) has inhibitory effects on the PI3K/Akt pathway. The authors therefore hypothesized that wounding would downregulate PTEN and that this downregulation would enhance wound healing.METHODS: In human corneal epithelial (HCE) cell monolayer and rat cornea scratch wound models, the authors investigated PTEN and Akt expression using Western blot and immunofluorescence analyses. The effects of PTEN and PI3K inhibitors dipotassium bisperoxo (picolinato) oxovanadate (bpv(pic)) and LY294002 on cell migration and wound closure were investigated using time-lapse imaging. Finally, the authors investigated the effect of PTEN inhibition on wound healing in whole rat eyes.RESULTS: In HCE cell monolayer and rat cornea, PTEN was downregulated at the wound edges within 30 minutes of wounding. The downregulation of PTEN was causal in a simultaneous increase in Akt activation, which was responsible for a significant increase in individual cell migration rate from 8.8 μm/h to 17.3 μm/h. An increased migration rate was maintained for 20 hours. PTEN inhibition significantly enhanced the wound healing rate in the HCE cell monolayer from 10 minutes onward after treatment and reduced the healing time in eye organ culture from 30 to 20 hours.CONCLUSIONS: Injury to the corneal epithelium downregulates the expression of PTEN at wound edges, allowing increased PI3K/Akt signaling, thereby contributing to a significant enhancement of cell migration and wound healing. These results suggest that PTEN inhibition may be an effective treatment for corneal injury.

KW - animals

KW - blotting, Western

KW - cell movement

KW - chromones

KW - down-regulation

KW - enzyme inhibitors

KW - epithelium, corneal

KW - female

KW - fluorescent antibody technique, indirect

KW - humans

KW - male

KW - morpholines

KW - organ culture techniques

KW - organometallic compounds

KW - PTEN phosphohydrolase

KW - phosphatidylinositol 3-kinases

KW - proto-oncogene proteins c-akt

KW - rats

KW - rats, sprague-dawley

KW - wound healing

U2 - 10.1167/iovs.10-5972

DO - 10.1167/iovs.10-5972

M3 - Article

VL - 52

SP - 2272

EP - 2278

JO - Investigative Ophthalmology & Visual Science

JF - Investigative Ophthalmology & Visual Science

SN - 0146-0404

IS - 5

ER -