Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011 Jun 6;88(23-24):1047-54.
doi: 10.1016/j.lfs.2011.03.016. Epub 2011 Apr 3.

Resveratrol in combination with other dietary polyphenols concomitantly enhances antiproliferation and UGT1A1 induction in Caco-2 cells

Otito F Iwuchukwu  1 , Ronald J Tallarida, Swati Nagar
Affiliations

Resveratrol in combination with other dietary polyphenols concomitantly enhances antiproliferation and UGT1A1 induction in Caco-2 cells

Otito F Iwuchukwu et al. Life Sci. .

Abstract

Aims: The only FDA approved medication for colorectal cancer (CRC) prevention is celecoxib. Its adverse effects underline the need for safer drugs. Polyphenols like resveratrol are in clinical trials for this purpose. This study aimed at examining effects of resveratrol alone and in combination with curcumin or chrysin on UGT induction in Caco-2 cells. Phytochemical combinations were selected using drug combination analyses of various anti-proliferation ratios of resveratrol+curcumin and resveratrol+chrysin.

Main methods: Cell proliferation and UGT1A1 induction assays were carried out with individual polyphenols and combinations. Cell viability was determined with AlamarBlue assays. UGT1A1 mRNA was quantified via real time RT-PCR. UGT activity was determined with 4-methylumbelliferone (4MU) glucuronidation.

Key findings: Cell proliferation IC(50) estimates (± SE) for resveratrol, curcumin and chrysin were 20.8 ± 1.2, 20.1 ± 1.1 and 16.3 ± 1.3μM respectively. Combination of anti-proliferative effects showed additivity for resveratrol+chrysin and resveratrol+curcumin. Resveratrol at its IC(50) mediated a four-fold induction of UGT1A1 mRNA in a concentration independent manner. Chrysin at its IC(50) induced UGT1A1 expression seven-fold while Curcumin at its IC(90) mediated a two-fold induction. The 20 μM:40μ M resveratrol+curcumin and 20 μM :32 μM resveratrol+chrysin combinations mediated the greatest increases in mRNA expression (12 and 22 folds respectively). Significant increase in 4-MU glucuronidation was observed with combinations exhibiting maximal mRNA induction.

Significance: Phytochemical combinations can offer greater chemoprevention than single agents. These chemicals might offer safer options than present synthetic therapeutics for CRC prevention.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Caco-2 cell cytotoxicity dose response curves for 1 – 100 μM resveratrol (RES), curcumin (Curc) and chrysin. Data are expressed as Mean ± SE, n = 4. Representative fitted lines are depicted; IC50 estimates were obtained by fitting the model to actual data replicates.
Figure 2
Figure 2
Induction of UGT 1A1 mRNA in post confluent Caco-2 cells treated with varying concentrations of RES. Data are expressed as mean + SD, n = 6. ***significantly different from control (p < 0.001).
Figure 3
Figure 3
Studies with RES + chrysin combinations. A) Caco-2 cell cytotoxicity observed upon 72 h treatment with IC50 combinations of RES + chrysin. RES + chrysin 1:1 is 20 uM RES + 16 uM chrysin, 1:2 is 20 uM RES + 32 uM chrysin, and 2:1 is 40 uM RES + 16 uM chrysin. Data expressed as mean + SD, n = 4 *** greater than RES alone (p < 0.001); +++ greater than either RES alone and chrysin alone (p < 0.001) using a one way ANOVA and post hoc Tukey’s multiple comparison analysis. B) Dose-effect data for RES and chrysin plotted as the magnitude of the percent reduction in growth against log dose. C) Induction of UGT 1A1 mRNA in post confluent Caco-2 cells treated for 72h with IC50 combinations of RES + chrysin. Data are expressed as mean + SD, n = 3.*** Groups significantly different from control (p < 0.001). +++different from RES alone or chrysin alone (p< 0.001) using a one way ANOVA followed by a post hoc Student-Newman-Keuls multiple comparison analysis.
Figure 3
Figure 3
Studies with RES + chrysin combinations. A) Caco-2 cell cytotoxicity observed upon 72 h treatment with IC50 combinations of RES + chrysin. RES + chrysin 1:1 is 20 uM RES + 16 uM chrysin, 1:2 is 20 uM RES + 32 uM chrysin, and 2:1 is 40 uM RES + 16 uM chrysin. Data expressed as mean + SD, n = 4 *** greater than RES alone (p < 0.001); +++ greater than either RES alone and chrysin alone (p < 0.001) using a one way ANOVA and post hoc Tukey’s multiple comparison analysis. B) Dose-effect data for RES and chrysin plotted as the magnitude of the percent reduction in growth against log dose. C) Induction of UGT 1A1 mRNA in post confluent Caco-2 cells treated for 72h with IC50 combinations of RES + chrysin. Data are expressed as mean + SD, n = 3.*** Groups significantly different from control (p < 0.001). +++different from RES alone or chrysin alone (p< 0.001) using a one way ANOVA followed by a post hoc Student-Newman-Keuls multiple comparison analysis.
Figure 4
Figure 4
Studies with RES + Curc combinations. A) Caco-2 cell growth inhibition observed upon 72 h treatment with IC50 combinations of RES + Curc where RES + Curc 1:1 is 20 uM RES + 20 uM Curc, 1:2 is 20 uM RES + 40 uM Curc, and 2:1 is 40 uM RES + 20 uM Curc. *** greater than RES alone and Curcumin alone (p < 0.001); +++ greater than 1:1 RES + Curc and 2:1 Res + Curc (p < 0.001) using a one way ANOVA and post hoc Tukey’s multiple comparison analysis. B) Cell growth inhibition observed with RES + Curc IC90 combinations where RES IC50 and Curc IC90 concentrations were used to determine ratios and RES + Curc 1:1 is 20 uM RES + 40 uM Curc, 1:2 is 20 uM RES + 80 uM Curc, and 2:1 is 40 uM RES + 40 uM Curc. Data expressed as mean + SD, n = 4. *** greater than RES alone (p < 0.001). C) Induction of UGT 1A1 mRNA in post confluent Caco-2 cells treated for 72h with RES IC50 + Curc IC90 combinations. ***significantly different from control (p < 0.001) * (p < 0.05), +++different from RES alone and Curc alone (p< 0.001) N.D = not detectable
Figure 4
Figure 4
Studies with RES + Curc combinations. A) Caco-2 cell growth inhibition observed upon 72 h treatment with IC50 combinations of RES + Curc where RES + Curc 1:1 is 20 uM RES + 20 uM Curc, 1:2 is 20 uM RES + 40 uM Curc, and 2:1 is 40 uM RES + 20 uM Curc. *** greater than RES alone and Curcumin alone (p < 0.001); +++ greater than 1:1 RES + Curc and 2:1 Res + Curc (p < 0.001) using a one way ANOVA and post hoc Tukey’s multiple comparison analysis. B) Cell growth inhibition observed with RES + Curc IC90 combinations where RES IC50 and Curc IC90 concentrations were used to determine ratios and RES + Curc 1:1 is 20 uM RES + 40 uM Curc, 1:2 is 20 uM RES + 80 uM Curc, and 2:1 is 40 uM RES + 40 uM Curc. Data expressed as mean + SD, n = 4. *** greater than RES alone (p < 0.001). C) Induction of UGT 1A1 mRNA in post confluent Caco-2 cells treated for 72h with RES IC50 + Curc IC90 combinations. ***significantly different from control (p < 0.001) * (p < 0.05), +++different from RES alone and Curc alone (p< 0.001) N.D = not detectable
Figure 5
Figure 5
Michaelis-Menten curves for 4-MUG formation in control and treated Caco-2 whole cell homogenates. 4-MU glucuronidation assays were conducted using a substrate concentration range of 0-1000uM. Experiments were conducted in triplicate and data are expressed as Mean ± SE. Representative fitted lines are depicted; kinetics estimates were obtained by fitting the model to actual data replicates.
See this image and copyright information in PMC

References

    1. Goswami SK, Das DK. Resveratrol and chemoprevention. Cancer Lett. 2009;284(1):1–6. - PubMed
    1. Kundu JK, Surh YJ. Cancer chemopreventive and therapeutic potential of resveratrol: mechanistic perspectives. Cancer Lett. 2008;269(2):243–61. - PubMed
    1. Hebbar V, Shen G, Hu R, Kim BR, Chen C, Korytko PJ, Crowell JA, Levine BS, Kong AN. Toxicogenomics of resveratrol in rat liver. Life Sci. 2005;76(20):2299–314. - PubMed
    1. Walle T, Ta N, Kawamori T, Wen X, Tsuji PA, Walle UK. Cancer chemopreventive properties of orally bioavailable flavonoids--methylated versus unmethylated flavones. Biochem Pharmacol. 2007;73(9):1288–96. - PMC - PubMed
    1. Szaefer H, Cichocki M, Brauze D, Baer-Dubowska W. Alteration in phase I and II enzyme activities and polycyclic aromatic hydrocarbons-DNA adduct formation by plant phenolics in mouse epidermis. Nutr Cancer. 2004;48(1):70–7. - PubMed

Publication types

MeSH terms