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1 Acta Neurobiologiae Experimentalis

1 2009

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Defferential modulation of basal and IFN-induced stat target genes expression by specific stat directed oligodeoxynucleotide decoys in glioma C6 cells

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Adach-Kilon A. , Dabrowski M. , Kaminska B.

nr 3

Constitutive activation of STAT3, leading to up-regulation of some proliferation-related and anti-apoptotic genes, was described in a number of tumors, including glioblastomas and is considered as an important drug target for cancer therapy. The binding sites for STAT3 are highly similar to that of STAT1, and the two factors may have opposite effects on expression of the same target genes. We took advantage of differences in binding sequences, relatively specifi c for some but not all STAT proteins, to elucidate the interplay between different STATs in the regulation of gene expression in the rat glioma C6 cells. A number of double-stranded oligodeoxynucleotide (ODN) decoys, each carrying a particular STAT binding sequence, were employed as competitive inhibitors of binding sites in the genome. We compared the effects of the decoy containing ISRE/GAS (binding complexes of STAT1, STAT2, IRFs) or STAT1 site (specifi c for STAT1 homodimer) on the expression of endogenous genes. We report that the decoy against STAT1 reduced the constitutive expression of endogenous STAT3 target genes under basal conditions. The decoy ISRE/GAS blocked IFNg-induced increase in expression of STAT target genes. Decoys with mutated and/or scrambled STAT-binding motifs were used as controls for specifi city. Our results demonstrate a limited usefulness of some ODN decoys in manipulation of endogenous gene expression, probably due to limited sequence specifi city of STAT binding sites.