Title

Reduction of DNA damage by interaction s of dietary compounds with DNA.

Poster Number

5

Format

Poster Presentation

Abstract/Artist Statement

Plant-derived dietary compounds and supplements are potential sources of chemicals that reduce cancer risk. The purpose of this study was to examine the effect of genistein, a soy-derived isoflavone, on DNA adduct formation. Cisplatin, a DNA damaging agent used in anti-cancer treatment, was selected as a model for assaying genotoxicity in vitro. Since DNA damage frequently occurs at "hotspot" sequences, we chose the human p53 gene (exon 7 - 9) as "hotspot" target DNA. Target and excess of carrier DNA were incubated with cisplatin in the presence of genistein or control chemicals overnight at 37°C in buffered saline solution. The extent of DNA damage was detected by a polymerase chain reaction (PCR) and agarose gel electrophoresis assay developed for this purpose. Reduction in the amount of a p53-specific PCR product was taken as evidence of DNA damage. Genistein was found to negatively affect cisplatin-induced DNA damage. A significant retention of band intensity was observed at 2 mM genistein. Quercetin, another plant-derived flavone had a similar effect at the same concentration. The intercalator ethidium bromide also restored band intensity, indicating intercalation could be a mechanism involved in DNA damage reduction by flavones and isoflavones.

Location

Pacific Geosciences Center

Start Date

26-4-2003 9:00 AM

End Date

26-4-2003 5:00 PM

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Apr 26th, 9:00 AM Apr 26th, 5:00 PM

Reduction of DNA damage by interaction s of dietary compounds with DNA.

Pacific Geosciences Center

Plant-derived dietary compounds and supplements are potential sources of chemicals that reduce cancer risk. The purpose of this study was to examine the effect of genistein, a soy-derived isoflavone, on DNA adduct formation. Cisplatin, a DNA damaging agent used in anti-cancer treatment, was selected as a model for assaying genotoxicity in vitro. Since DNA damage frequently occurs at "hotspot" sequences, we chose the human p53 gene (exon 7 - 9) as "hotspot" target DNA. Target and excess of carrier DNA were incubated with cisplatin in the presence of genistein or control chemicals overnight at 37°C in buffered saline solution. The extent of DNA damage was detected by a polymerase chain reaction (PCR) and agarose gel electrophoresis assay developed for this purpose. Reduction in the amount of a p53-specific PCR product was taken as evidence of DNA damage. Genistein was found to negatively affect cisplatin-induced DNA damage. A significant retention of band intensity was observed at 2 mM genistein. Quercetin, another plant-derived flavone had a similar effect at the same concentration. The intercalator ethidium bromide also restored band intensity, indicating intercalation could be a mechanism involved in DNA damage reduction by flavones and isoflavones.