Isofraxidin, a Coumarin Component from Acanthopanax senticosus, Inhibits Matrix Metalloproteinase-7 Expression and Cell Invasion of Human Hepatoma Cells

Abstract

Invasion and metastasis of cancer cells are the major causes of high motality in cancer patients. Matrix metalloproteinases (MMPs) are zinc-dependent proteolytic enzymes capable of degrading the components of extracellular matrix (ECM), including type IV collagen, gelatin and fibronectin. Cancer cells destruct surrounding tissues by enzymatic action of MMPs, leading to cancer invasion and metastasis. Among several MMPs, MMP-2, -7 and -9 are considered to be involved in cancer invasion and metastasis. In particular, MMP-7 is frequently found to overexpress in the invasive edge of cancer cells and play important roles in invasion or metastasis of digestive cancer. 1,2) Recently, the attempt to search for MMP inhibitors from plants is being performed, and several investigators have reported the inhibitory effects of epigallocatechin, 3) caffeic acid, 4) and proanthocyanin 5) on gene expression or enzymatic activity of MMP-2 and -9. However, except for the papers by Kawabata et al.,[6][7][8] little attention has been given to the development of MMP-7 inhibitors.Isofraxidin occurs in a variety of plants including Acansopanax senticosus HARMs (Japanese name: Ezoukogi). As the major compounds of Acanthopanax senticosus, eleutheroside B, B1, E, isofraxidin, and chlorogenic acid have been identified, isolated, and reported to possess several biological activities, such as anti-stress, anti-fatigue, antigastric ulcer, anti-depressive, immuno-enhancing and antiinflammatory effects. [9][10][11][12][13][14][15][16] We previously examined the effect of isofraxidin on the expression of inflammatory mediator cyclooxygenase-2 (COX-2) mRNA and found inhibitory effects at a significant level on human synovial sarcoma cell line SW982 cells. [11][12][13][14][15][16] In this study, we investigated isofraxidin in its anti-tumor effects on human hepatoma cell lines in cellular and molecular terms. MATERIALS AND METHODSCell Culture Human hepatoma cell lines HuH-7, HepG2 and HLE and a human lung fibroblast cell line IMR90 were obtained from Japanese Collection of Research Bioresources (JCRB). Cells were grown in RPMI-1640 culture medium supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 mg/ml streptomycin at 37°C in a humidified atmosphere of 5% CO 2 and 95% air.Phenolic Compounds Isofraxidin was purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Eleutheroside E was purchased from Phytochem Referenzsubstanzen GbRmb (Ichenhausen, Germany). The compounds were added to the culture medium at appropriate concentrations as a dimethylsulfoxide (DMSO) solution (the final DMSO concentration was 0.5% in all cultures), and the same amount of vehicle was added to the control cultures.3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium Bromide (MTT) Assay For cell attachment study, cells were plated in 96-well plates at 3.0ϫ10 4 cells/cm 2 (low cell density) in 0.1 ml of serum-free RPMI-1640 medium containing or not containing isofraxidin and incubated for 24 h. For cell growth study, cells were plated i...

Activity-guided isolation and identification of the major antioxidant and anticancer compounds from a commercial Pelargonium sidoides tincture

“…GA and proanthocyanidins have been shown to arrest the cell cycle, induce apoptosis, inhibit angiogenesis and tumor invasion in vitro (Ji et al, 2009;Ho et al, 2010;Madlener et al, 2007;Nandakumar et al, 2008). Anti-proliferative activity has also been reported for simple coumarins like 7-hydroxycoumarin (LopezGonzalez et al, 2004;Elinos-Báez et al, 2005), esculetin (Chu et al, 2001;Lee et al, 2008), 4-hydroxycoumarin (Velasco-Velázquez et al, 2003isofraxidin (Borris et al, 1980;Yamazaki and Tokiwa, 2010) and 7,8-dihydroxy-4-methyl coumarin (Riveiro et al, 2008). These coumarins were observed to arrest the cell cycle, induce apoptosis and inhibit tumor cell migration and invasion.…”

Isofraxidin exhibited anti-inflammatory effects in vivo and inhibited TNF-α production in LPS-induced mouse peritoneal macrophages in vitro via the MAPK pathway

“…In addition, it acts at the transcription level to regulate inflammatory modulators, such as IκBα and NF-κB, that could affect downstream genes involved in the inflammatory response, including iNOS, COX-2, and TNF-α [22]. However, many studies have shown that IF did not suppress COX-2 gene expression pronounced [20,25,26]. It was also reported that IF showed an inhibitory effect on the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in hepatoma cells, but it showed little effect on activator protein-1 (AP-1) DNA binding activity, nuclear factor-kappa B (NF-κB) nuclear translocation, and inhibitory kappa B (IκB) degradation [26].…”

“…However, many studies have shown that IF did not suppress COX-2 gene expression pronounced [20,25,26]. It was also reported that IF showed an inhibitory effect on the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in hepatoma cells, but it showed little effect on activator protein-1 (AP-1) DNA binding activity, nuclear factor-kappa B (NF-κB) nuclear translocation, and inhibitory kappa B (IκB) degradation [26]. All these findings strongly suggest that IF may have a significant effect on inflammation and play an important role in the MAPK pathway.…”

The novel secretory protein CGREF1 inhibits the activation of AP-1 transcriptional activity and cell proliferation

“…proliferation and transformation, and is associated with the invasion and metastasis of cancer cells. Therefore, the ERK MAPK signaling pathway is one of the most important targets for the development of anti-cancer drugs (Yamazaki and Tokiwa, 2010;Johnson et al, 2010). Recent studies have demonstrated that ERK1/2 and p38 regulate trophoblasts differentiation in human term placenta (Daoud et al, 2005), and the ERK signaling pathway may be essential for regulating the balance between proliferation and apoptosis in CD34 + -derived neutrophil progenitor cells (Geest et al, 2009).…”