Elsevier

Biomedicine & Pharmacotherapy

Volume 70, March 2015, Pages 140-145
Biomedicine & Pharmacotherapy

Original article
Effects of natural nuclear factor-kappa B inhibitors on anticancer drug efflux transporter human P-glycoprotein

https://doi.org/10.1016/j.biopha.2015.01.007Get rights and content

Abstract

Drug efflux transporter P-glycoprotein plays an important role in cancer chemotherapy. The nuclear factor-κB (NF-κB) transcription factors play critical roles in development and progression of cancer. In this study, the effects of natural compounds that can inhibit NF-κB activation on the function of P-glycoprotein were investigated using human MDR1 gene-transfected KB/MDR1 cells. The accumulation of daunorubicin or rhodamine 123, fluorescent substrates of P-glycoprotein, in KB/MDR1 cells increased in the presence of caffeic acid phenetyl ester (CAPE), licochalcone A, anacardic acid, celastrol, xanthohumol, magnolol, and honokiol in a concentration-dependent manner. In contrast, lupeol, zerumbone, thymoquinone, emodin, and anethol had no effects. The ATPase activities of P-glycoprotein were stimulated by CAPE, licochalcone A, anacardic acid, celastrol, xanthohumol, magnolol, and honokiol. Tumor necrosis factor (TNF)-α stimulated NF-κB activation was inhibited by CAPE, licochalcone A, anacardic acid, and xanthohumol. KB/MDR1 cells were sensitized to vinblastine cytotoxicity by CAPE, licochalcone A, anacardic acid, xanthohumol, magnolol, and honokiol, showing that these natural NF-κB inhibitors reverse multidrug resistance. These results suggest that natural compounds, such as CAPE, licochalcone A, and anacardic acid, have dual inhibitory effects on the anticancer drug efflux transporter P-glycoprotein and NF-κB activation, and may become useful to enhance the efficacy of cancer chemotherapy.

Introduction

Multidrug resistance is a phenomenon of resistance of tumors to a wide spectrum of anticancer drugs, and is the significant obstacle to the success of cancer chemotherapy. Multidrug resistance can have many causes, but most commonly encountered mechanism is the increased efflux of cytotoxic drugs by energy-dependent transporters [1], [2], [3]. P-glycoprotein belongs to the large ATP-binding cassette (ABC) transporter superfamily of membrane transport proteins, and is the first member to be identified [4]. The MDR1 (ABCB1) gene is located on chromosome 7q21. It consists of 28 exons, which encode a 1280-amino acid glycoprotein. P-glycoprotein mediates resistance to a variety of pharmacologically unrelated anticancer drugs, such as vinblastine, daunorubicin, doxorubicin, epirubicin, etoposide, imatinib, irinotecan, and paclitaxel [1], [2], [3]. We previously reported that natural compounds including flavonoids, such as quercetin and (-)-epigallocatechin gallate (EGCG), curcumin, capsaicin, [6]-gingerol, auraptene, and guggulsterone, could reverse the multidrug resistance by the inhibition of anticancer drug efflux transporter P-glycoprotein [5], [6], [7], [8], [9], [10].

The signaling pathways that govern cell proliferation, survival and oncogenesis are of prime interest in the biology of cancer. The nuclear factor-κB (NF-κB) transcription factors (p50/p105 (NF-κB1), p52/p100 (NF-κB2), RelA (p65), RelB, c-Rel) and the pathways that control NF-κB activation play critical roles in the regulation of inflammation and oncogenesis [11], [12], [13]. In most resting cells, NF-κB is sequestered in the cytoplasm by binding to the inhibitory IκB proteins. NF-κB is activated by a variety of stimuli such as carcinogens, inflammatory agents including phorbol esters, bacterial endotoxins, and tumor necrosis factor (TNF). These stimuli promote dissociation of IκB through phosphorylation by the IκB kinase (IKK), ubiquitination and proteasome-mediated degradation. This process unmasks the nuclear localization sequences of NF-κB, which then accumulates in the nucleus, binds to κB regulatory elements and activates target genes including critical anti-apoptotic proteins, such as IAPs (inhibitor of apoptosis proteins) and Bcl-2 [11], [12], [13]. The genes encoding anti-apoptotic proteins function as oncogenes, activation of NF-κB promote tumor development. In many cancers, NF-κB is constitutively activated by mutations in genes that control NF-κB activation [11], [12], [13].

Fruits and vegetables are excellent sources of fiber, vitamins, and minerals, but they also contain various natural compounds that may provide substantial health benefits beyond basic nutrition [14], [15], [16]. Epidemiological and experimental studies have highlighted the ability of dietary natural compounds to reduce the risk of cancer [14], [15], [16]. Several dietary phytochemicals, such as EGCG, curcumin, capsaicin, guggulsterone, caffeic acid phenetyl ester (CAPE), emodin, and anethol are known to block the NF-κB activation process, and thus act as cancer chemopreventive agents [14], [15], [16].

Natural compounds that can inhibit both of the NF-κB activation and anticancer drug efflux transporter would be beneficial for enhancing the efficacy of cancer chemotherapy. In this study, we investigated the interaction of natural compound that is reported to inhibit NF-κB activation with human P-glycoprotein.

Section snippets

Materials

Dulbecco's modified Eagle's medium (D-MEM) and fetal bovine serum (FBS) were purchased from Life Technologies (Carlsbad, CA). Vinblastine, daunorubicin, rhodamine 123, TNF-α, zerumbone, eugenol, magnolol, and honokiol were obtained from Wako Pure Chemical Industries, Ltd (Osaka, Japan). Withaferin A, ganoderic acid A, and xanthohumol were obtained from Tokiwa Phytochemical Co, Ltd (Chiba, Japan). Celastrol and maslinic acid were from Cayman Chemical (Ann Arbor, MI). Anacardic acid was purchased

Effects of natural compounds on the accumulation of daunorubicin in KB/MDR1 cells

Fig. 1 shows the chemical structures of several natural compounds used in this study. We first tested the accumulation of daunorubicin in human P-glycoprotein-overexpressing KB/MDR1 cells in the presence of 50 μM natural compounds (Fig. 2). CAPE, licochalcone A, anacardic acid, celastrol, xanthohumol, magnolol, and honokiol increased the cellular accumulation of daunorubicin, which indicates that these compounds inhibit the P-glycoprotein-mediated efflux of daunorubicin. In contrast, lupeol,

Discussion

Cancer chemotherapy is usually a marginal proposition in the sense that the maximum dose tolerated by the patient is often barely sufficient to kill a useful percentage of the cancer cells. Relatively small increases in drug resistance in cancer cells are thus sufficient to render the drug ineffective. P-glycoprotein is expressed at cancer cell membranes and can cause multidrug resistance. Therefore, P-glycoprotein seems to be a good target for circumventing multidrug resistance. Since the

Acknowledgements

We thank Prof. Kazumitsu Ueda of Kyoto University for providing KB/MDR1 cells. This work was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (Grant Number 22590151) (T.N.).

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    Chemical compounds studied in this article: caffeic acid phenetyl ester (PubChem CID: 5281787); licochalcone A (PubChem CID: 5318998); anacardic acid (PubChem CID: 167551); celastrol (PubChem CID: 122724); xanthohumol (PubChem CID: 639665); magnolol (PubChem CID: 72300); honokiol (PubChem CID: 72303).

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