Reaction of keratinocytes to exogenous DNA.
Keratinocytes have the ability to take up oligonucleotides (ODN) and plasmid DNA probably by receptor-mediated endocytosis or macropinocytosis. Despite the use of DNA for antisense and gene therapy, little is known about the regulation of genes following exposure to nucleic acids. To systematically identify gene regulation in keratinocytes upon exposure to ODN, we screened human cytokine DNA arrays containing 383 different genes and found interleukin-1 alpha (IL-1 alpha), IL-1 beta, integrin beta1, alpha-tubulin and follistatin greatly induced, while most genes were unaffected. The time course and concentration dependence for IL-1 alpha and follistatin was confirmed by the standard Northern blot technique and found to be induced by picomolar or femtomolar concentrations of ODN. ODN of different length and sequence induced comparable amounts of IL-1 alpha and follistatin. Their induction was independent of the negative charge and of several proinflammatory compounds and proteins such as lipopolysaccharides, IL-1 beta or IFN-gamma, but was partly inhibited by activin A. In summary, our study revealed several genes of the acute phase protein family that were induced in a non-sequence-specific manner following the exposure of normal human keratinocytes to ODN. Therefore, it is tempting to speculate that, upon internalization, ODN bind to an intracellular receptor (e.g. Toll-like receptor 9), which mediates signaling.
Mirmohammadsadegh
Alireza
A
Department of Dermatology, Heinrich Heine University, Düsseldorf, Germany. a.sadegh@uni-duesseldorf.de
Maschke
Jan
J
Basner-Tschakarjan
Etiena
E
Bär
Annett
A
Hengge
U lrich R
Ul
D028341:Activins / Q000494:pharmacology; D001483:Base Sequence / Q000235:genetics; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005260:Female; D038681:Follistatin / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D005820:Genetic Testing; D006497:Heparan Sulfate / Q000494:pharmacology; D006801:Humans; D018836:Inflammation Mediators / Q000494:pharmacology; D028322:Inhibin-beta Subunits / Q000494:pharmacology; D019012:Integrin beta1 / Q000378:metabolism; D007375:Interleukin-1 / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000378:metabolism / Q000494:pharmacology*; D011930:Reaction Time / Q000187:drug effects / Q000235:genetics; D014404:Tubulin / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects* / Q000235:genetics
D028341:Activins; D004247:DNA; D038681:Follistatin; D006497:Heparan Sulfate; D018836:Inflammation Mediators; D028322:Inhibin-beta Subunits; D019012:Integrin beta1; D007375:Interleukin-1; D009841:Oligonucleotides; D014404:Tubulin; C422405:activin A