Cell lines and transgenic mice expressing a matrix metalloproteinase-9 promoter-driven reporter gene: potential for assay of ultraviolet light effects and light-inhibiting compounds.
Acute and chronic exposure to ultraviolet (UV) wavelengths in sunlight can cause adverse reactions in exposed areas of the skin and corneas. UV exposure up-regulates the synthesis of Matrix Metalloproteinses (MMPs) and evidence suggests these enzymes mediate tissue damage. Therefore MMP gene activity can serve as a surrogate marker for bioassays. In this study, we tested the possible utility for this purpose of two stably transfected cell lines (from mouse keratinocytes and rabbit epithelial-like corneal cells) and a transgenic mouse line (line 3445), each harboring a DNA construct containing a bacterial beta-galactosidase (LacZ) reporter gene driven by the rabbit MMP-9 transcriptional promoter. We observed only a weak 2-fold maximal induction of LacZ reporter gene expression in the mouse epidermal cell line after exposure to UV-B irradiation (5, 10, 40 mJ/cm2) and no significant expression of the reporter gene in the rabbit epithelial-like cell line. Similarly negative results were obtained when primary corneal epithelial cells from human and rabbit were exposed to different doses of UV-B irradiation and endogenous MMP-9 gene expression was assayed by zymography and immunoprecipitation analysis. In contrast, when skin from 3-day-old transgenic mouse line 3445 was exposed to UV-B and UV-A, a clear dose-dependent induction of the LacZ reporter gene occurred and the location of gene expression was dependent on the wave-length of irradiation. These results suggest that line 3445 transgenic mice may serve as a useful tool to quantitatively and qualitatively assess the biological effects of UV light and the efficacy of therapeutic agents.
Bargagna-Mohan
Paola
P
Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, USA.
Mohan
Royce
R
Russo
Laoti
L
Kochevar
Irene E
IE
Fini
M Elizabeth
ME
D000818:Animals; D002460:Cell Line; D019573:Epithelium, Corneal / Q000528:radiation effects; D005786:Gene Expression Regulation / Q000528:radiation effects*; D017930:Genes, Reporter / Q000235:genetics*; D007763:Lac Operon / Q000235:genetics / Q000502:physiology; D020780:Matrix Metalloproteinase 9 / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011817:Rabbits; D012867:Skin / Q000528:radiation effects; D014466:Ultraviolet Rays
D020780:Matrix Metalloproteinase 9