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.
Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, USA.
Paola
P
Bargagna-Mohan
Royce
R
Mohan
Laoti
L
Russo
Irene E
IE
Kochevar
M Elizabeth
ME
Fini
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