RNA fingerprinting displays UVB-specific disruption of transcriptional control in human melanocytes.
In mammalian cells, UV induces a limited set of early transcribed genes, which overlaps with the set of genes induced by tumor promoting drugs such as 12-O-tetradecanoyl phorbol-13-acetate (TPA). Among these are genes for transcription factors, the activation of which leads to complex secondary changes in expression of multiple target genes. How these delayed pleiotropic UV effects on transcription may contribute to initiation of melanoma skin cancer is poorly understood. We analyzed changes in the relative abundances of 1900 transcripts in newborn human melanocytes 8 h after treatment with UVB, TPA, and cycloheximide in all combinations, using RNA arbitrarily primed PCR for differential display. The relative abundances of 205 transcripts (11% of all transcripts surveyed) were altered by one or more of the treatment combinations. Fourteen of the 77 genes up-regulated by TPA were also up-regulated by UVB, but 60 of the TPA up-regulated genes were down-regulated by UVB, indicating both intersecting and independent signal transduction pathways for UVB and TPA. A number of UVB and TPA target genes were identified by cDNA cloning. Consistent with UVB induction of a partly transformed phenotype in mammalian cells, UVB antagonized the TPA-inducible expression of tumor-suppressive tropomyosin 3 mRNA. In addition, UVB may impair mitochondrial functioning and induce oxidative stress by strong down-regulation of mitochondrial transcription. Finally, increased expression of the dihydropteridine reductase gene, a major regulator of the cellular tetrahydrobiopterin pool, was linked to the UV pathway.
Vogt
T M
TM
Sidney Kimmel Cancer Center, San Diego, California 92121, USA.
Welsh
J
J
Stolz
W
W
Kullmann
F
F
Jung
B
B
Landthaler
M
M
McClelland
M
M
D002273:Carcinogens / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects / Q000528:radiation effects; D003513:Cycloheximide / Q000494:pharmacology; D016172:DNA Fingerprinting; D006801:Humans; D008544:Melanocytes / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012313:RNA / Q000187:drug effects / Q000528:radiation effects*; D012333:RNA, Messenger / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects / Q000528:radiation effects*; D014466:Ultraviolet Rays
D002273:Carcinogens; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D012313:RNA; D003513:Cycloheximide; D013755:Tetradecanoylphorbol Acetate