Effects of UV, 4-NQO and TPA on gene expression in cultured human epidermal keratinocytes

• Published in: Nucleic acids research Vol. 15; no. 15; pp. 5945 - 5962
• Main Authors: KARTASOVA, T, CORNELISSEN, B. J. C, BELT, P, VAN DE PUTTE, P
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 11.08.1987
• Subjects: 4-Nitroquinoline-1-oxide - pharmacology; Amino Acid Sequence; Base Sequence; Biological and medical sciences; Cells, Cultured; Cysteine Proteinase Inhibitors; DNA - isolation & purification; Epidermis - drug effects; Epidermis - metabolism; Epidermis - radiation effects; Fundamental and applied biological sciences. Psychology; Gene expression; Humans; Keratins - genetics; Molecular and cellular biology; Molecular genetics; Nitroquinolines - pharmacology; Nucleic Acid Hybridization; Protease Inhibitors - genetics; RNA, Messenger - biosynthesis; RNA, Messenger - drug effects; RNA, Messenger - radiation effects; Tetradecanoylphorbol Acetate - pharmacology; Human origin; Cell culture; Keratinocyte; Ultraviolet radiation; Gene expression; Mechanism of action
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/15.15.5945

In an approach to study effects of UV light on gene expression in human epidermal keratinocytes, a cDNA library was constructed from poly(A)RNA isolated after UV irradiation from cultured keratinocytes. The cDNA library was differentially screened with labelled cDNA probes synthesized on poly(A)RNA isolated from UV irradiated or nonirradiated keratinocytes. Forty clones were selected and subjected to further analysis, 31 of them are described in this report. Whereas total mRNA synthesis is reduced after UV irradiation or treatment with 4-NQO Northern blot analysis revealed that there is an at least relative increase in the level of mRNAs corresponding to the majority of the isolated cDNA clones. Among these 15 were identified as corresponding to mRNAs for 50K and 56K keratins and for 50K- and 46K-related keratin. In addition, clones were found corresponding to the proteinase inhibitor cystatin A and to the glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Treatment of keratinocytes with the tumor promoter TPA had no effect on the mRNA level for most of the clones except those corresponding to keratins. Our results indicate that in keratinocytes UV irradiation leads to a relative increase in the level of some mRNAs.