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Nucleic Acids Research Advance Access published online on April 22, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp252
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© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Oligonucleotides suppress PKB/Akt and act as superinductors of apoptosis in human keratinocytes

Stefan Kippenberger1,*, Jutta Müller1, Maike Schultz1, Annette Dorn1, Andreas Bock2, Hüseyin Aygün3, Diamant Thaçi1, Matthias Hofmann1, Roland Kaufmann1 and August Bernd1

1Department of Dermatology and Venerology, J.W. Goethe-University, 60590 Frankfurt/Main, 2Henkel AG & Co. KGaA, Henkelstr. 67, 40191 Düsseldorf and 3BioSpring GmbH, Alt Fechenheim 34, 60386 Frankfurt, Germany

*To whom correspondence should be addressed. Tel: +49 69 6301 7734; Fax: +49 69 6301 6466; Email: kippenberger{at}em.uni-frankfurt.de

Received January 28, 2009. Revised April 3, 2009. Accepted April 3, 2009.

DNA oligonucleotides (ODN) applied to an organism are known to modulate the innate and adaptive immune system. Previous studies showed that a CpG-containing ODN (CpG-1-PTO) and interestingly, also a non-CpG-containing ODN (nCpG-5-PTO) suppress inflammatory markers in skin. In the present study it was investigated whether these molecules also influence cell apoptosis. Here we show that CpG-1-PTO, nCpG-5-PTO, and also natural DNA suppress the phosphorylation of PKB/Akt in a cell-type-specific manner. Interestingly, only epithelial cells of the skin (normal human keratinocytes, HaCaT and A-431) show a suppression of PKB/Akt. This suppressive effect depends from ODN lengths, sequence and backbone. Moreover, it was found that TGF{alpha}-induced levels of PKB/Akt and EGFR were suppressed by the ODN tested. We hypothesize that this suppression might facilitate programmed cell death. By testing this hypothesis we found an increase of apoptosis markers (caspase 3/7, 8, 9, cytosolic cytochrome c, histone associated DNA fragments, apoptotic bodies) when cells were treated with ODN in combination with low doses of staurosporin, a well-known pro-apoptotic stimulus. In summary the present data demonstrate DNA as a modulator of apoptosis which specifically targets skin epithelial cells.


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