Genomic scale analysis of the human keratinocyte response to broad-band ultraviolet-B irradiation.
Ultraviolet B (UVB) radiation is an important inducer of many biologic changes in skin, of which keratinocytes are a key target. To gain better insight into changes in gene expression generated in the early phase after UVB exposure, we used complementary RNA (cRNA) microarray hybridization to compare differences in mRNA expression of UVB-irradiated (single dose of 100 J/m2 broad-band UVB) and sham-irradiated primary cultured human keratinocytes. Six hours after irradiation, total RNA was isolated from keratinocytes, and cRNA was synthesized and hybridized to a GeneChip expression array (Affymetrix) consisting of 6800 genes. Based on a threshold of > twofold change, 187 genes (2.8%) were designated to be the most UVB-responsive. Surprisingly, none of these genes had been shown previously to be modulated by UVB. Conversely, several genes in the microarray that had been reported previously to be UVB- responsive by other methods showed less (< twofold) or no change. Northern blotting of seven differentially modulated genes produced results similar to those derived from microarray technology, thereby validating the accuracy of screening. Clustering based on known or likely functions indicated that among 88 upregulated genes, nine encode for cytochrome c subunits, six for ribosomal proteins, and two for regulators of apoptosis. By contrast, many of the 99 downregulated genes are involved in transcription, differentiation and transport. These findings indicate that keratinocytes respond to a single low dose of broad-band UVB irradiation by enhancing processes involved in energy production and translation, while suppressing those related to transcription, differentiation and transport.
Takao
Joe
J
Department of Dermatology, The University of Texas South-western Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9069, USA.
Ariizumi
Kiyoshi
K
Dougherty
Irene I
II
Cruz
Ponciano D
PD
D017209:Apoptosis / Q000235:genetics; D015152:Blotting, Northern; D002478:Cells, Cultured; D003574:Cytochrome c Group / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000528:radiation effects*; D020411:Oligonucleotide Array Sequence Analysis; D018075:RNA, Complementary / Q000378:metabolism; D011829:Radiation Dosage; D012269:Ribosomal Proteins / Q000235:genetics; D014466:Ultraviolet Rays
D003574:Cytochrome c Group; D018075:RNA, Complementary; D012269:Ribosomal Proteins