Elsevier

Biochemical Pharmacology

Volume 70, Issue 5, 1 September 2005, Pages 700-713
Biochemical Pharmacology

Curcumin (diferuloylmethane) inhibits constitutive NF-κB activation, induces G1/S arrest, suppresses proliferation, and induces apoptosis in mantle cell lymphoma

https://doi.org/10.1016/j.bcp.2005.04.043Get rights and content

Abstract

Human mantle cell lymphoma (MCL), an aggressive B cell non-Hodgkin's lymphoma, is characterized by the overexpression of cyclin D1 which plays an essential role in the survival and proliferation of MCL. Because of MCL's resistance to current chemotherapy, novel approaches are needed. Since MCL cells are known to overexpress NF-κB regulated gene products (including cyclin D1), we used curcumin, a pharmacologically safe agent, to target NF-κB in a variety of MCL cell lines. All four MCL cell lines examined had overexpression of cyclin D1, constitutive active NF-κB and IκB kinase and phosphorylated forms of IκBα and p65. This correlated with expression of TNF, IκBα, Bcl-2, Bcl-xl, COX-2 and IL-6, all regulated by NF-κB. On treatment of cells with curcumin, however, downregulated constitutive active NF-κB and inhibited the consitutively active IκBα kinase (IKK), and phosphorylation of IκBα and p65. Curcumin also inhibited constitutive activation of Akt, needed for IKK activation. Consequently, the expression of all NF-κB-regulated gene products, were downregulated by the polyphenol leading to the suppression of proliferation, cell cycle arrest at the G1/S phase of the cell cycle and induction of apoptosis as indicated by caspase activation, PARP cleavage, and annexin V staining. That NF-κB activation is directly linked to the proliferation of cells, is also indicated by the observation that peptide derived from the IKK/NEMO-binding domain and p65 suppressed the constitutive active NF-κB complex and inhibited the proliferation of MCL cells. Constitutive NF-κB activation was found to be due to TNF, as anti-TNF antibodies inhibited both NF-κB activation and proliferation of cells. Overall, our results indicate that curcumin inhibits the constitutive NF-κB and IKK leading to suppression of expression of NF-κB-regulated gene products that results in the suppression of proliferation, cell cycle arrest, and induction of apoptosis in MCL.

Introduction

Mantle cell lymphoma (MCL) is a type of B cell non-Hodgkin lymphoma (NHL) that accounts for 3–10% of all NHL in Western countries [1] and leads to higher fraction of deaths, given that it is an incurable malignancy [2]. MCL patients are most often elderly men who present with advance stage of disease, and most often with extranodal involvement [3]. The length of survival of MCL patients following diagnosis is quite variable with median survival of around 3 years. Depending upon the severity, it may vary between 1 and 10 years.

There is no clear standard approach for treating mantle cell lymphoma. Chemotherapy with chlorambucil [4], CVP (cyclophosphamide, vincristine, and prednisone), or CHOP (cyclophosphamide, hydroxydoxorubicin, oncovin, and prednisone) [5] can be used in patients who are not candidates for aggressive therapy. Allogeneic transplant is promising for young patients with matched donors [6]. High-dose therapy with autologous stem cell transplantation may extend the time to progression and the duration of overall survival. Other agents, including rituximab [7], fludarabine [8], and cladribine [9] have demonstrated activity, but these agents do not appear to offer survival advantages over combination chemotherapy. Despite these treatment options, this malignancy remains incurable.

MCL is characterized cytogenetically by the presence of a non-random chromosomal abnormality, the t(11;14)(q13;q32) chromosomal translocation, as a result of which the cyclin D1 gene is brought under the control of the immunoglobulin heavy chain gene enhancer, leading to overexpression of cyclin D1 [10], [11], [12]. The latter is a hallmark of this disease and is believed to contribute to deregulated cellular proliferation in MCL [2]. Additionally, the anti-apoptotic protein Bcl2 is also overexpressed in MCL [13]. Recent gene profiling studies have shown that the genes involved in TNF and NF-κB signaling pathways are overexpressed in MCL [14]. Both cyclin D1 and Bcl-2 are regulated by NF-κB [15], [16].

NF-κB is a transcription factor present in the cytoplasm as an inactive heterotrimer consisting of p50, p65, and IκBα subunits. On activation, IκBα undergoes phosphorylation and ubiquitination-dependent degradation leading to nuclear translocation and binding to a specific consensus sequence in the DNA which results in gene transcription [17]. The kinase which phosphorylates IκBα is termed IκB kinase (IKK) composed of IKKα, IKKβ and IKKγ (also called NEMO [18]). NF-κB regulates the expression of genes involved in antiapoptosis (e.g. bcl-2 and bcl-xl); proliferation (COX-2 and cyclin D1) and metastasis (e.g., MMP-9).

Curcumin, a diferuloylmethane derived from turmeric (Curcuma longa) is a pharmacologically safe agent that has been shown to suppress NF-κB activation and NF-κB gene products [19], [20]. In the current report, we targeted NF-κB pathway in MCL cells by using curcumin. We found that all four MCL cell lines expressed constitutively active NF-κB and NF-κB-regulated gene products (Bcl-2, Bcl-XL, cyclin D1, COX-2, TNF, IL-6, RANK, and RANKL); and treatment with curcumin suppressed NF-κB activation and downregulated the expression of these gene products leading to cell cycle arrest, suppression of proliferation and induction of apoptosis.

Section snippets

Materials

The four MCL cell lines included in this study were JeKo-1, Mino, SP-53, and Granta 519. JeKo-1 [21] was kindly provided by T. Akagi (Chosun University Medical School, Kwangju, Korea). The cell line Mino was established and characterized at The University of Texas M. D. Anderson Cancer Center by Dr. Raymond Lai [22]. SP-53 [23] was a generous gift from M. Daibata (Kochi Medical School, Kochi, Japan). The cell line Granta 519 was purchased from Deutsche Sammlung von Mikroorganismen und

Results

The aim of this study was to investigate the effect of curcumin on the proliferation and survival of MCL. Curcumin was selected because it is a pharmacologically safe agent that has been shown to downregulate cyclin D1 expression (for references see [20]). Four MCL cell lines that have been previously characterized were used in the present study [29]. As the MCL cells are characterized by overexpression of cyclin D1, so we also examined the expression of cyclin D1 in MCL cells. All four MCL

Discussion

In this study, we examined the effect of curcumin on the constitutive NF-κB activation in MCL cells. Our results show that NF-κB is constitutively expressed in all the four MCL cell lines that we examined. All these cells showed constitutive activation of IKK, constitutive phosphorylation of p65, and IκBα and overexpression of mRNAs for TNF, IL-6, RANK and RANKL. MCL cells also overexpressed IκBα, Bcl-2, Bcl-XL and COX-2 that are known to be regulated by NF-κB. Treatment of MCL cells with

Acknowledgments

We would like to thank Walter Pagel for a careful review of the manuscript. Dr. Aggarwal is a Ransom Horne Jr. Distinguished Professor of Cancer Research. This work was supported in part by the Odyssey Program and the Theodore N. Law Award for Scientific Achievement at The University of Texas M. D. Anderson Cancer Center (to SS).

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  • Cited by (0)

    Supported by the Clayton Foundation for Research (to BBA), a Department of Defense US Army Breast Cancer Research Program grant (BC010610, to BBA), a PO1 grant (CA91844) from the National Institutes of Health on lung chemoprevention (to BBA).

    1

    Present address: Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alta., Canada.

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