Molecular and Cellular Pharmacology Demethoxycurcumin suppresses migration and invasion of MDA-MB-231 human breast cancer cell line Supachai Yodkeeree a,c , Chadarat Ampasavate b , Bokyung Sung c , Bharat B. Aggarwal c , Pornngarm Limtrakul a, ⁎ a Deparment of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand b Deparment of Pharmaceutical Science, Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand c Cytokine Research Laboratory, Department of Experimental Therapeutics, The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030, USA abstract article info Article history: Received 15 May 2009 Received in revised form 11 September 2009 Accepted 28 September 2009 Available online 7 October 2009 Keywords: Curcuminoids Demethoxycurcumin Matrix metalloproteinases (MMPs) Urokinase plasminogen activator (uPA) Nuclear factor-kappa B (NF-κB) Cancer invasion Demethoxycurcumin (DMC) is one of the main active compounds of curcuminoids found in turmeric powder, which is used as a spice in Asian cooking and traditional medicine. Recent studies reveal that DMC has several biological activities including anti-inflammation and anti-cancer activities. However, the molecular mechanism by which DMC has anti-metastasis activity in breast cancer cells remains poorly understood. Here, we report for the first time that DMC inhibited adhesion, migration and invasion of MDA-MB-231 human breast cancer cells. For cancer cell migration and invasion, extracellular matrix (ECM) degradation processes are required. MDA-MB-231 cells treated with DMC had decreased levels of ECM degradation-associated proteins including matrix metalloproteinase-9 (MMP-9), membrane type-1 matrix metalloproteinase (MT1-MMP), urokinase plasmino- gen activator (uPA) and uPA receptor (uPAR), while the level of uPA inhibitor (PAI-1) was up-regulated. Moreover, DMC also reduced the expression of intercellular adhesion molecule-1 (ICAM-1) and chemokine receptor 4, (CXCR4), which is involved in modulation of the tumor metastasis process. We also found that DMC treatment inhibited the DNA binding activity of nuclear factor-kappa B (NF-κB), which is known to mediate the expression of MMPs, uPA, uPAR, ICAM-1, and CXCR4. These findings strongly suggest that the mechanism of DMC- mediated anti-invasive activity involves modulation of the expression of invasion-associated proteins, possibly by targeting NF-κB in MDA-MB-231 cells. © 2009 Elsevier B.V. All rights reserved. 1. Introduction Cancer invasion and metastasis are the leading causes of mortality in patients with breast cancer. Tumor metastasis consists of a complex cascade of events, including cell adhesion, invasion, and angiogenesis (Cavallaro and Christofori, 2001). Degradation of the extracellular matrix (ECM) by proteolytic enzymes is a crucial step in tumor metastasis. Among these enzymes, matrix metalloproteinase (MMPs) and urokinase plasminogen activator (uPA) play a key role in degrading components of the ECM (Liotta et al., 1980). Many reports have demonstrated that uPA plays an importance role in tumor metastasis and over-expression of uPA in breast cancer is a strong indicator of a poor prognosis (Bertrand et al., 2002; Look et al., 2002). The uPA system consists of uPA, its receptor uPAR and its inhibitor, plasminogen inhibitor type 1 and type 2 (PAI-1 and -2). Upon binding of uPA and uPAR, active uPA converts plasminogen to the active serine protease plasmin (Dano et al., 1999), which is involved in the degradation of the ECM. Moreover, the binding of uPA to uPAR also generates signal transduction that allows enhanced cell migration (Tang and Wei, 2008). MMPs are a multigene family of zinc- dependent endopeptidases capable of degrading essentially all ECM components. In breast cancer, up-regulation of several MMPs, such as MMP-2, -3, -7, -13 and MT1-MMP, is generally associated with breast cancer metastasis (Balduyck et al., 2000; Mylona et al., 2007). Therefore, to regulate the expression of ECM degradation enzymes are considered as a target for therapeutic intervention. Nuclear factor κB (NF-κB), a transcription factor, plays an important role in carcinogenesis as well as in the regulation of inflammatory responses. Several genes involved in tumor metastasis have been identified as being regulated by NF-κB. A recent study reported that NF-κB promotes migration and metastasis of breast cancer cells through up-regulation of the chemokine receptor, CXCR4 (Helbig et al., 2003), and adhesive molecule, ICAM-1 (Lerebours et al., 2008). Moreover, over- expression of MMPs and uPA is also regulated by NF-κB(Connelly et al., 2007; Sliva et al., 2002). The frequent over-expression of NF-κB in tumor cells suggests that selected tumor cells may acquire metastatic activity by aberrant expression of metastasis relevant genes during their progression. Demethoxycurcumin (DMC) is a natural phenolic compound found in the rhizome of many Curcuma species such as, C. longa linn, C. aromatica and C. phaeocaulis (Zhang et al., 2008a). Generally, the rhizome of these species is utilized as a spice, flavoring agent, and in European Journal of Pharmacology 627 (2010) 8–15 ⁎ Corresponding author. Tel.: +66 053 945323; fax: +66 053 894031. E-mail address: plimtrak@mail.med.cmu.ac.th (P. Limtrakul). 0014-2999/$ – see front matter © 2009 Elsevier B.V. All rights reserved. doi:10.1016/j.ejphar.2009.09.052 Contents lists available at ScienceDirect European Journal of Pharmacology journal homepage: www.elsevier.com/locate/ejphar