Impact of Polyphenol Metabolites Produced by Colonic Microbiota on Expression of COX-2 and GSTT2 in Human Colon Cells (LT97)

Abstract

Polyphenols may play an important role in colon cancer prevention. After entering the colon, they are subjected to metabolism by the human gut microbiota. The objective of the present study was to analyze the impact of selected intestinal metabolites on modulation of enzymes involved in detoxification and inflammation in human adenoma cells LT97. LT97 cells were incubated with 3,4-dihydroxyphenylacetic acid (ES) and 3-(3,4-dihydroxyphenyl)-propionic acid (PS), metabolites of quercetin and chlorogenic acid/caffeic acid, respectively. The effect on cell number was analyzed using 4'- 6-diamino-2-phenylindole-dihydrochloride (DAPI)-staining. Modulation of glutathione S-transferase T2 (GSTT2) and cyclooxygenase-2 (COX-2) was measured by real-time PCR and Western blot. Comet assay was performed to assess the impact on DNA damage caused by the GSTT2 substrate cumene hydroperoxide (CumOOH). Polyphenol metabolites did not affect cell number but significantly upregulated GSTT2 expression and decreased COX-2. The latter was confirmed via Western blot. CumOOH-induced DNA damage was significantly reduced compared to the control. An upregulation of GSTT2 and downregulation of COX-2 could possibly contribute to the chemopreventive potential of polyphenols after degradation in the gut. Working with polyphenol metabolites is an important prerequisite to better understand the in vivo effects of pure polyphenols.

ACKNOWLEDGMENTS

The authors acknowledge grant funding from the Bundesministerium für Bildung und Forschung (FKZ 01EA503), Germany, and they thank Prof. Brigitte Marian, Institute of Cancer Research, University of Vienna, Austria for the generous gift of LT97 adenoma cells. Furthermore, we are grateful to PD Dr. Thomas Liehr for the kind provision of reagents and FISH probes to realize karyotyping of the LT97 cells.