ULTRAVIOLET ACTION SPECTRA FOR PHOTOBIOLOGICAL EFFECTS IN CULTURED HUMAN LENS EPITHELIAL CELLS

ULTRAVIOLET ACTION SPECTRA FOR PHOTOBIOLOGICAL EFFECTS IN CULTURED HUMAN LENS EPITHELIAL CELLS Abstract— The action spectrum for cell killing by UV radiation in human lens epithelial (HLE) cells is not known. Here we report the action spectrum in the 297–365 nm region in cultured HLE cells with an extended lifespan (HLE B‐3 cells) and define their usefulness as a model system for photobiological studies. Cells were irradiated with monochromatic radiation at 297, 302, 313, 325, 334 and 365 nm. Cell survival was determined using a clonogenic assay. Analysis of survival curves showed that radiation at 297 nm was six times more effective in cell killing than 302 nm radiation; 297 nm radiation was more than 260, 590, 1400 and 3000 times as effective in cell killing as 313, 325, 334 and 365 nm radiation, respectively. The action spectrum was similar in shape to that for other human epithelial cell lines and rabbit lens epithelial cells. The effect of UV radiation on crystallin synthesis was also determined at different wavelengths. To determine whether exposure to UV radiation affects the synthesis of β‐crystallin, cells were exposed to sublethal fluences of UV radiation at 302 and 313 nm, labeled with (35S)methionine and the newly synthesized βY‐crystallin was analyzed by immunoprecipitation and western blotting using an antibody to β‐crystallin. The results show a decrease in crystallin synthesis in HLE cells irradiated at 302 and 313 nm at fluences causing low cytotoxicity. The effect of radiation on membrane perturbation was determined by measuring enhancement of synthesis of prostaglandin E2 (PGE2). Synthesis of PGE2 occurs at all UV wavelengths tested in the 297–365 nm region. The slope of the PGE2 response curves was higher than that of cell killing curves in cultured HLE cells. These data show that cultured HLE cells with extended lifespan are a suitable system for investigating photobiological responses of cells to UV radiation. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Photochemistry & Photobiology Wiley

ULTRAVIOLET ACTION SPECTRA FOR PHOTOBIOLOGICAL EFFECTS IN CULTURED HUMAN LENS EPITHELIAL CELLS

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Publisher
Wiley
Copyright
Copyright © 1995 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0031-8655
eISSN
1751-1097
D.O.I.
10.1111/j.1751-1097.1995.tb09145.x
Publisher site
See Article on Publisher Site

Abstract

Abstract— The action spectrum for cell killing by UV radiation in human lens epithelial (HLE) cells is not known. Here we report the action spectrum in the 297–365 nm region in cultured HLE cells with an extended lifespan (HLE B‐3 cells) and define their usefulness as a model system for photobiological studies. Cells were irradiated with monochromatic radiation at 297, 302, 313, 325, 334 and 365 nm. Cell survival was determined using a clonogenic assay. Analysis of survival curves showed that radiation at 297 nm was six times more effective in cell killing than 302 nm radiation; 297 nm radiation was more than 260, 590, 1400 and 3000 times as effective in cell killing as 313, 325, 334 and 365 nm radiation, respectively. The action spectrum was similar in shape to that for other human epithelial cell lines and rabbit lens epithelial cells. The effect of UV radiation on crystallin synthesis was also determined at different wavelengths. To determine whether exposure to UV radiation affects the synthesis of β‐crystallin, cells were exposed to sublethal fluences of UV radiation at 302 and 313 nm, labeled with (35S)methionine and the newly synthesized βY‐crystallin was analyzed by immunoprecipitation and western blotting using an antibody to β‐crystallin. The results show a decrease in crystallin synthesis in HLE cells irradiated at 302 and 313 nm at fluences causing low cytotoxicity. The effect of radiation on membrane perturbation was determined by measuring enhancement of synthesis of prostaglandin E2 (PGE2). Synthesis of PGE2 occurs at all UV wavelengths tested in the 297–365 nm region. The slope of the PGE2 response curves was higher than that of cell killing curves in cultured HLE cells. These data show that cultured HLE cells with extended lifespan are a suitable system for investigating photobiological responses of cells to UV radiation.

Journal

Photochemistry & PhotobiologyWiley

Published: Nov 1, 1995

References

  • Comparative effects of UVA and UVB on cultured rabbit lens
    Hightower, Hightower; McCready, McCready
  • Membrane damage in UV‐irradiated lenses
    Hightower, Hightower; McCready, McCready; Borchman, Borchman
  • The effects of ultraviolet wavelengths of radiation present in sunlight on human cells in vitro
    Coohill, Coohill; Peak, Peak; Peak, Peak
  • Non‐nuclear damage and cell lysis are induced by UVA, but not UVB or UVC, radiation in three strains of L5178Y cells
    Beer, Beer; Olvey, Olvey; Miller, Miller; Thomas, Thomas; Godar, Godar
  • Enhanced keratinocyte prostaglandin synthesis after UV injury is due to increased phospholipase activity
    Kang‐Rotondo, Kang‐Rotondo; Miller, Miller; Morrison, Morrison; Pentland, Pentland

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