Impact of apple polyphenols on GSTT2 gene expression, subsequent protection of DNA and modulation of proliferation using LT97 human colon adenoma cells.
Apple extract (AE) enhances expression of glutathione S-transferases (e.g., GSTT2) in human colon cells (LT97). Therefore, aim of the present study was to identify functional consequences of GSTT2 induction by AE and to determine the relation of AE effects to isolated compounds. Polyphenol composition of AE was analyzed. LT97 cells were treated with AE or synthetic polyphenol mixture (SPM) under conditions that induced GSTT2, and challenged with GSTT2-2 substrate cumene hydroperoxide (CumOOH) to determine DNA damage using comet assay. Modulation of GSTT2 expression (real-time PCR) was reassessed, and the influence on cell proliferation and pro-oxidative potential of AE and SPM were assessed to understand additional mechanisms. Induction of GSTT2 by AE was accompanied by protection of LT97 cells from CumOOH-induced genotoxicity. Although SPM was unable to reflect AE-specific bioactivity related to GSTT2 modulation and anti-genotoxicity, inhibition of LT97 cell proliferation by SPM was comparable. Storage of AE caused changes in phenolic composition along with loss of activity regarding GSTT2 induction and amplified growth inhibition. At the applied concentrations, no H(2)O(2) formation was detectable with any of the substances. AE can protect against oxidatively induced DNA damage. Nevertheless, chemopreventive effects of AE strongly depend on the specific composition, which is modified by storage.
Miene
Claudia
C
Institute for Nutrition, Department of Nutritional Toxicology, Friedrich-Schiller-University Jena, Jena, Germany. claudia.miene@uni-jena.de
Klenow
Stefanie
S
Veeriah
Selvaraju
S
Richling
Elke
E
Glei
Michael
M
D000975:Antioxidants / Q000494:pharmacology*; D001555:Benzene Derivatives / Q000633:toxicity; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D020552:Comet Assay; D004249:DNA Damage; D005419:Flavonoids / Q000032:analysis / Q000494:pharmacology*; D005638:Fruit / Q000737:chemistry; D015971:Gene Expression Regulation, Enzymologic; D055785:Gene Knockdown Techniques; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis / Q000633:toxicity; D027845:Malus / Q000737:chemistry*; D016877:Oxidants / Q000633:toxicity; D010636:Phenols / Q000032:analysis / Q000494:pharmacology*; D010936:Plant Extracts / Q000494:pharmacology*; D059808:Polyphenols; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000975:Antioxidants; D001555:Benzene Derivatives; D005419:Flavonoids; D016877:Oxidants; D010636:Phenols; D010936:Plant Extracts; D059808:Polyphenols; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D006861:Hydrogen Peroxide; C491107:GSTT2 protein, human; D005982:Glutathione Transferase; C007164:cumene hydroperoxide