Effect of Curcumin on the Aryl Hydrocarbon Receptor and Cytochrome P450 1A1 in MCF-7 Human Breast Carcinoma Cells

doi:10.1016/S0006-2952(98)00143-9

Biochemical Pharmacology

Volume 56, Issue 2, 15 July 1998, Pages 197-206

https://doi.org/10.1016/S0006-2952(98)00143-9 Get rights and content

Abstract

We examined the interaction of curcumin, a dietary constituent and chemopreventive compound, with the carcinogen activation pathway mediated by the aryl hydrocarbon receptor (AhR) in MCF-7 mammary epithelial carcinoma cells. Curcumin caused a rapid accumulation of cytochrome P450 1A1 (CYP1A1) mRNA in a time- and concentration-dependent manner, and CYP1A1 monooxygenase activity increased as measured by ethoxyresorufin-O-deethylation. Curcumin activated the DNA-binding capacity of the AhR for the xenobiotic responsive element of CYP1A1 as measured by the electrophoretic-mobility shift assay (EMSA). Curcumin was able to compete with the prototypical AhR ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin for binding to the AhR in isolated MCF-7 cytosol, indicating that it interacts directly with the receptor. Although curcumin could activate the AhR on its own, it partially inhibited the activation of AhR, as measured by EMSA, and partially decreased the accumulation of CYP1A1 mRNA caused by the mammary carcinogen dimethylbenzanthracene (DMBA). Curcumin competitively inhibited CYP1A1 activity in DMBA-treated cells and in microsomes isolated from DMBA-treated cells. Curcumin also inhibited the metabolic activation of DMBA, as measured by the formation of DMBA–DNA adducts, and decreased DMBA-induced cytotoxicity. These results suggest that the chemopreventive effect of curcumin may be due, in part, to its ability to compete with aryl hydrocarbons for both the AhR and CYP1A1. Curcumin may thus be a natural ligand and substrate of the AhR pathway.

Section snippets

Materials

RPMI 1640, glutamine, fetal bovine serum, trypsin/EDTA, PBS, and TBE buffer were obtained from BioFluids. Curcumin, DMBA, HEPES, EDTA, dithiothreitol (DTT), glycerol, poly(dI-dC), sodium molybdate, ethyoxyresorufin, resorufin, Tris–HCl, sulforhodamine, salmon sperm, and protease inhibitors were from the Sigma Chemical Co. α-Naphthoflavone was from Indofine. [³²P]CTP and [³²P]dATP were from DuPont/NEN. The RT–PCR kit was from Stratagene. TBE gels, TBE running buffer, and high-density sample

Effect of Curcumin on CYP1A1 mRNA

We measured CYP1A1 mRNA in MCF-7 cells treated with DMBA in the presence or absence of curcumin using RT–PCR. Treatment of cells with 1 μM of DMBA for 24 hr caused a 15-fold increase in CYP1A1 mRNA accumulation (Fig. 1). Curcumin at 5 μM of partially inhibited the DMBA-induced increase. Surprisingly, treatment with curcumin alone caused an approximately six-fold increase in CYP1A1 mRNA. We therefore examined the effect of curcumin in the absence of other treatment on CYP1A1 mRNA. Treatment

Discussion

We examined the effect of curcumin on the AhR and the major carcinogen-activating enzyme in MCF-7 cells, CYP1A1 [25]. Using RT–PCR, we found that the mammary carcinogen DMBA caused an increase in CYP1A1 mRNA in MCF-7 cells that was partially antagonized by simultaneous treatment of the cells with curcumin (Fig. 1). It has been shown previously that the ability of the liganded AhR to induce transcription of genes depends on its ability to bind enhancer sequences, called the XRE, flanking the

Acknowledgements

The authors would like to thank Dr. Miriam Poirier for her critical reading of the manuscript.

References (32)

P Limtrakul et al.
Inhibitory effect of dietary curcumin on skin carcinogenesis in mice
Cancer Lett
(1997)
K Singletary et al.
Inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumorigenesis and DMBA–DNA adduct formation by curcumin
Cancer Lett
(1996)
S Oetari et al.
Effects of curcumin on cytochrome P450 and glutathione S-transferase activities in rat liver
Biochem Pharmacol
(1996)
SS Deshpande et al.
Effects of curcumin on the formation of benzo[a]pyrene derived DNA adducts in vitro
Cancer Lett
(1995)
PF Firozi et al.
Action of curcumin on the cytochrome P450-system catalyzing the activation of aflatoxin B₁
Chem Biol Interact
(1996)
O Dohr et al.
Different response of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-sensitive genes in human breast cancer MCF-7 and MDA-MB 231 cells
Arch Biochem Biophys
(1995)
M Moore et al.
Benzo[a]pyrene-resistant MCF-7 human breast cancer cells. A unique aryl hydrocarbon-nonresponsive clone
J Biol Chem
(1994)
P Chomczynski et al.
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction
Anal Biochem
(1987)
MS Denison et al.
The DNA recognition site for the dioxin–Ah receptor complexNucleotide sequence and functional analysis
J Biol Chem
(1988)
Y-H Chen et al.
Protein kinase C modulates regulation of the CYP1A1 gene by the aryl hydrocarbon receptor
J Biol Chem
(1996)

L Poellinger et al.

A hydroxylapatite microassay for receptor binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin and 3-methylcholanthrene in various target tissues

Anal Biochem

(1985)

MM Bradford

A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding

Anal Biochem

(1976)

SW Kennedy et al.

Simultaneous measurement of cytochrome P4501A catalytic activity and total protein concentration with a fluorescence plate reader

Anal Biochem

(1994)

OP Sharma

Antioxidant activity of curcumin and related compounds

Biochem Pharmacol

(1976)

ML Kuo et al.

Curcumin, an antioxidant and anti-tumor promoter, induces apoptosis in human leukemia cells

Biochim Biophys Acta

(1996)

N Dzeletovic et al.

Regulation of dioxin receptor function by omeprazole

J Biol Chem

(1997)

Cited by (228)

Targeting aryl hydrocarbon receptor to prevent cancer in barrier organs
2024, Biochemical Pharmacology
The skin, lung, and gut are important barrier organs that control how the body reacts to environmental stressors such as ultraviolet (UV) radiation, air pollutants, dietary components, and microorganisms. The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that plays an important role in maintaining homeostasis of barrier organs. AhR was initially discovered as a receptor for environmental chemical carcinogens such as polycyclic aromatic hydrocarbons (PAHs). Activation of AhR pathways by PAHs leads to increased DNA damage and mutations which ultimately lead to carcinogenesis. Ongoing evidence reveals an ever-expanding role of AhR. Recently, AhR has been linked to immune systems by the interaction with the development of natural killer (NK) cells, regulatory T (T_reg) cells, and T helper 17 (Th17) cells, as well as the production of immunosuppressive cytokines. However, the role of AhR in carcinogenesis is not as straightforward as we initially thought. Although AhR activation has been shown to promote carcinogenesis in some studies, others suggest that it may act as a tumor suppressor. In this review, we aim to explore the role of AhR in the development of cancer that originates from barrier organs. We also examined the preclinical efficacy data of AhR agonists and antagonists on carcinogenesis to determine whether AhR modulation can be a viable option for cancer chemoprevention.
Functions of the aryl hydrocarbon receptor (AHR) beyond the canonical AHR/ARNT signaling pathway
2023, Biochemical Pharmacology
The aryl hydrocarbon receptor (AHR) is a ligand-dependent transcription factor regulating adaptive and maladaptive responses toward exogenous and endogenous signals. Research from various biomedical disciplines has provided compelling evidence that the AHR is critically involved in the pathogenesis of a variety of diseases and disorders, including autoimmunity, inflammatory diseases, endocrine disruption, premature aging and cancer. Accordingly, AHR is considered an attractive target for the development of novel preventive and therapeutic measures. However, the ligand-based targeting of AHR is considerably complicated by the fact that the receptor does not always follow the beaten track, i.e. the canonical AHR/ARNT signaling pathway. Instead, AHR might team up with other transcription factors and signaling molecules to shape gene expression patterns and associated physiological or pathophysiological functions in a ligand-, cell- and micromilieu-dependent manner. Herein, we provide an overview about some of the most important non-canonical functions of AHR, including crosstalk with major signaling pathways involved in controlling cell fate and function, immune responses, adaptation to low oxygen levels and oxidative stress, ubiquitination and proteasomal degradation. Further research on these diverse and exciting yet often ambivalent facets of AHR biology is urgently needed in order to exploit the full potential of AHR modulation for disease prevention and treatment.
Emodin inhibits U87 glioblastoma cells migration by activating aryl hydrocarbon receptor (AhR) signaling pathway
2022, Ecotoxicology and Environmental Safety
Aryl hydrocarbon receptor (AhR) is a ligand-activated receptor to mediates the biological reactions of many environmental and natural compounds, which is highly expressed in glioblastoma. Although it has been reported that AhR agonist emodin can suppress some kinds of tumors, its inhibitory effect on glioblastoma migration and its relationship with AhR remain unclear. Based on the complexity of tumor pathogenesis and the tissue specificity of AhR, we hope can further understand the effect of emodin on glioblastoma and explore its mechanism. We found that the inhibitory effect of emodin on the migration of U87 glioblastoma cells increased with time, and the cell migration ability was inhibited by about 25% after 36 h exposure. In this process, emodin promoted the expression of the tumor suppressor IL24 by activating the AhR signaling pathway. Reducing the expression of AhR or IL24 by interfering RNA could block or relieve the inhibitory effect of emodin on the U87 cells migration, which indicates the inhibition of emodin on the migration of glioblastoma is mediated by the AhR-IL24 axis. Our data proved the AhR-IL24 signal axis is an important pathway for emodin to inhibit the migration of glioblastoma, and the AhR signaling pathway can be used as a key target to research the regulation effect and its mechanism of compounds on glioblastoma migration.
Curcumin as a privileged scaffold molecule for various biological targets in drug development
2022, Studies in Natural Products Chemistry
Curcumin [1,7‐bis(4‐hydroxy‐3‐methoxyphenyl) ‐1,6‐heptadiene‐3,5‐dione] is pharmacologically active polyphenol isolated from rhizome Curcumin longa L and is a versatile molecule known for its various biological properties including anti-inflammatory, antioxidant, antibacterial, including antibacterial, anticancer, arthritis, allergies and other various chronic diseases. In recent years this compound has attracted considerable attention due to multimodal effects. In this endeavor, novel curcumin derivatives have also been synthesized and tested against different models of human disease. The current chapter is a compilation of existing information from many recent studies utilizing curcumin in various disease conditions and the new drug delivery approach to tackle bioavailability issues have also been discussed. Further the ongoing clinical studies on curcumin in various disease conditions are also reviewed in this chapter.
The ameliorative effect of curcumin on hepatic CYP1A1 and CYP1A2 genes dysregulation and hepatorenal damage induced by fenitrothion oral intoxication in male rats
2021, Pesticide Biochemistry and Physiology
This research aimed to assess curcumin (CUR) effects on fenitrothion (FNT), a broad-spectrum organophosphate insecticide, −induced hepatorenal damage. Thirty adult male Wistar rats were allocated at random to five equal groups orally administered distilled water containing 1% carboxyl methylcellulose, corn oil (1 mL/rat), CUR (100 mg/kg b.wt.), FNT (5 mg/kg b.wt.), or CUR + FNT. CUR and FNT were dosed three times a week for two months. At the end of this trial, blood and tissue samples (liver and kidney) were subjected to molecular, biochemical, and histopathological assessments. The results revealed that CUR significantly diminished the FNT-induced up-regulation of hepatic CYP1A1 and CYP1A2 transcriptional levels. Moreover, CUR significantly suppressed the increment of the serum levels of hepatic alanine aminotransferase, gamma-glutamyl transferase, and kidney damage indicators (urea and creatinine) in FNT-intoxicated rats. Furthermore, in the hepatic and renal tissues, CUR remarkably restored the FNT-associated depletion of the antioxidant enzymes (glutathione peroxidase, glutathione reductase, glutathione S transferase, catalase, and superoxide dismutase). In addition, CUR notably reduced the FNT-induced increment in malondialdehyde content in the hepatic and renal tissues. Besides, the pathological aberrations in liver and kidney tissues resulting from FNT exposure were significantly abolished in FNT + CUR treated rats. Overall, CUR could be an effective ameliorative agent against negative pesticide impacts like FNT.
Mapping multiple endocrine disrupting activities in Virginia rivers using effect-based assays
2021, Science of the Total Environment
Water sources are frequently contaminated with natural and anthropogenic substances having known or suspected endocrine disrupting activities; however, these activities are not routinely measured and monitored. Phenotypic bioassays are a promising new approach for detection and quantitation of endocrine disrupting chemicals (EDCs). We developed cell lines expressing fluorescent chimeric constructs capable of detecting environmental contaminants which interact with multiple nuclear receptors. Using these assays, we tested water samples collected in the summers of 2016, 2017 and 2018 from two major Virginia rivers. Samples were concentrated 200× and screened for contaminants interacting with the androgen (AR), glucocorticoid (GR), aryl hydrocarbon (AhR) and thyroid receptors. Among 45 tested sites, over 70% had AR activity and 60% had AhR activity. Many sites were also positive for GR and TRβ activation (22% and 42%, respectively). Multiple sites were positive for more than one type of contaminants, indicating presence of complex mixtures. These activities may negatively impact river ecosystems and consequently human health.

View all citing articles on Scopus

View full text

Original ArticlesEffect of Curcumin on the Aryl Hydrocarbon Receptor and Cytochrome P450 1A1 in MCF-7 Human Breast Carcinoma Cells

Abstract

Section snippets

Materials

Effect of Curcumin on CYP1A1 mRNA

Discussion

Acknowledgements

Cancer Lett

Cancer Lett

Biochem Pharmacol

Cancer Lett

Chem Biol Interact

Arch Biochem Biophys

J Biol Chem

Anal Biochem

J Biol Chem

J Biol Chem

Anal Biochem

Anal Biochem

Anal Biochem

Biochem Pharmacol

Biochim Biophys Acta

J Biol Chem

Original Articles
Effect of Curcumin on the Aryl Hydrocarbon Receptor and Cytochrome P450 1A1 in MCF-7 Human Breast Carcinoma Cells