liu.seSearch for publications in DiVA
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
UVB radiation increases EGF-R gene expression in cultured human normal melanocytes
Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Epidermal growth factor (EGF) and its receptor are thought to play important roles in the UV response of mammalian cells. Since UVB radiation has been shown to influence melanocyte growth and development, we sought to determine whether UVB would alter the expression of the gene for EGF-R. Using Northern blot, the present study exanlined the expression of EGF-R mRNA in cultured normal human melanocytes exposed to a single physiologic dose of UVB radiation. Total cellular RNA was extracted from cultured normal human melanocytes at various time-points after irradiation or sham-irradiation. Constitutive expression ofEGF-Rmfu'!A was detected in the melanocytes. After UVB irradiation with a single dose of 6o or 180 mJ/cm2, the EGF-R rnRNA decreased within 12 h to approximately 50 % of the initial value. After 24 h there was a subsequent increase and a peak at 36 h, showing a three-fold increase. After 48 hours a decline was seen, but the expression of EGF-R mRNA was still doubled as compared to control.

Using flow cytometric analysis EGF-R on melanocytes were detected. The median EGF-R immunofluorescence was decreased after UVB-irradiation as compared to non-irradiated cells. The lowest EGF-R immunofluorescence was seen 12 hours after UVB irradiation. Witi1in 48 hours post irradiation we could not detect any increase in EGF-R that we would correlate to the increased EGFR gene expression seen at 36 hours. We conclude ti1at the steady-state levels of EGF-R mRNAis upregulated by single exposure to UVB.

National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-81075OAI: oai:DiVA.org:liu-81075DiVA: diva2:550202
Available from: 2012-09-06 Created: 2012-09-06 Last updated: 2012-09-06Bibliographically approved
In thesis
1. Dynamics and gene expression of growth factor receptors in human cultured skin cells: Effects of UV radiation and calcium on EGF- and PDGF-receptors
Open this publication in new window or tab >>Dynamics and gene expression of growth factor receptors in human cultured skin cells: Effects of UV radiation and calcium on EGF- and PDGF-receptors
2001 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Growth factors transmit biological signals for the stimulation of cell growth and their stimulation may be involved in turnourigenesis. It is therefore of great importance to understand growth factor receptor reactions in response to stimuli such as calcium depletion or ultraviolet radiation, which normal human cells are invariably exposed to during their growth cycle. We have studied human skin cells i.e. fibroblasts, lceratinocytes and melanocytes and their growth factor receptor expression on the surface of cells, reactions in the plane of the cell membrane, intracellular trafficlcing, and gene expression after exposure to their ligand, serum, UVB radiation and calcium depletion. We have used Fluorescence recovery after photo bleaching (FRAP) to assess receptor characteristics in cell membranes, confocal laser scanning microscopy to visualize receptor internalization, Ratio imaging for calcium studies, Northern blot for detection of the gene for the epidermal growth factor receptor (EGF-R) and flow cytometry for cell surface receptor determination. Platelet-derived growth factor receptors (PDGF-R) and EGF-R were studied since they affect cell proliferation and viability.

Our results showed that the addition of PDGF increased receptor mobility characteristics in normal fibroblasts, also "starvation" of cells increased their receptor mobility. We could show that changes in both intra-and extracellular free Ca2+ influence the mobility characteristics of PDGF-ß2 receptors. We have demonstrated that the three celltypes display different basal EGF-R mobility characteristics. After UVB irradiation mobility characteristics increased in all cell types but with differences in diffusion coefficients or mobile fractions. Addition of antioxidant enzymes, catalase and superoxide dismutase, prior to UVB irradiated cells abolished the UY-induced receptor mobility changes. We have shown that a single physiologic dose ofUVB radiation alters the intracellular EGF-R distribution and intracellular transport in melanocytes. It also significantly alters the melanocyte phenotype. We were able to detect a constitutive EGF-R gene expression and showed that UVB-radiation induces a time-dependent induction in EGF-R mRNA in melanocytes. Human melanocytes express EGF-R on their cell surface and UVR induces time dependent changes in the number of receptors but the number of receptors does not correlate with the level of UV-induced EGF-R gene expression.

It is concluded that UV-radiation, growth factors and calcium, ubiquitous constituents of every day life, all having tremendous effects in vivo, also affect human cells in vitro in parameters studied that are of importance for proliferation, survival and tumourigenesis.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2001. 88 p.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 674
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25680 (URN)10056 (Local ID)91-7219-965-2 (ISBN)10056 (Archive number)10056 (OAI)
Public defence
2001-05-18, Administrationshusets Aula, Universitetssjukhuset, Linköping, 09:30 (Swedish)
Opponent
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2012-09-06Bibliographically approved

Open Access in DiVA

No full text

Authority records BETA

Hallbeck, Anna-LottaBriheim, KristinaWasteson, Åke

Search in DiVA

By author/editor
Hallbeck, Anna-LottaBriheim, KristinaWasteson, Åke
By organisation
Cell biologyFaculty of Health Sciences
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar

urn-nbn

Altmetric score

urn-nbn
Total: 48 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf