Gene profiling study of G3139- and Bcl-2-targeting siRNAs identifies a unique G3139 molecular signature
| Autor: | Luba Benimetskaya, Diane D. Ilsley, Anastasia Khvorova, Cy A. Stein, Paul S. Miller, Emily M. Anderson, William Marshall |
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| Rok vydání: | 2005 |
| Předmět: |
Male
Cancer Research Small interfering RNA Time Factors Biology Transfection Downregulation and upregulation Cell Line Tumor Humans Gene silencing Gene Silencing RNA Small Interfering Molecular Biology Gene Cell Proliferation Oligonucleotide Array Sequence Analysis Messenger RNA Oligonucleotide Gene Expression Profiling Prostatic Neoplasms Thionucleotides Molecular biology Gene Expression Regulation Neoplastic Proto-Oncogene Proteins c-bcl-2 Lipofectamine Molecular Medicine RNA Interference |
| Zdroj: | Cancer Gene Therapy. 13:406-414 |
| ISSN: | 1476-5500 0929-1903 |
| DOI: | 10.1038/sj.cgt.7700901 |
| Popis: | G3139 is a phosphorothioate oligodeoxyribonucleotide that is targeted to the initiation codon region of the Bcl-2 mRNA, which downregulates Bcl-2 protein and mRNA expression via an antisense mechanism. In previous work, we have demonstrated that the phenotype observed in several prostate and melanoma cell lines after treatment with G3139 appears to be Bcl-2 independent. In contrast, downregulation of Bcl-2 expression by a small interfering RNA (siRNA) produced little or no phenotype change. In the present work, we performed an Agilent oligonucleotide microarray assay on mRNA isolated from PC3 prostate cancer cells that were treated with two different oligonucleotide gene-silencing reagents. G3139 and a Bcl-2-targeting siRNA both downregulate Bcl-2 expression, but via different mechanisms. A side-by-side comparative analysis showed that the expression profile generated by these molecules differs substantially. The study revealed upregulation of the expression of stress-inducible genes in PC3 cells at 1 and 3 days after a 5-h transfection with G3139 complexed with Lipofectamine 2000. In contrast, only a very diminished stress response was seen 1 and 3 days after a 24-h transfection of siRNA/Lipofectamine 2000 complexes. These results highlight the profound differences in off-target effects in cells treated with the phosphorothioate oligonucleotide G3139 and with an siRNA targeted to the same gene, and provide further evidence that the mechanism of action of G3139 is not related to Bcl-2 silencing. |
| Databáze: | OpenAIRE |
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