Tissue Transglutaminase Regulates Matrix Metalloproteinase-2 in Ovarian Cancer by Modulating cAMP-response Element-binding Protein Activity

Indiana University Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.
Journal of Biological Chemistry (Impact Factor: 4.57). 04/2009; 284(23):15390-9. DOI: 10.1074/jbc.M808331200
Source: PubMed


Tissue transglutaminase 2 (TG2) is overexpressed in epithelial ovarian cancer (EOC) and promotes intraperitoneal metastasis. How TG2 facilitates the spread of EOC is unknown. Here, we show that TG2 regulates the expression and function of matrix metalloproteinase-2 (MMP-2), a critical mediator of tissue invasiveness. TG2 knockdown down-regulates MMP-2 protein and mRNA expression in SKOV3, IGROV-1, MDA-MB-436, and PC-3 cancer cells. TG2 knockdown or inhibition of TG2 activity using KCC009 decreases MMP-2 gelatinase activity in cancer cells. MMP-2 expression and function are regulated by TG2 at transcriptional level, as demonstrated by quantitative PCR and reporter assays. We used bioinformatics and chromatin immunoprecipitation to identify a CREB binding site in the MMP-2 promoter. Binding of CREB to the MMP-2 promoter was diminished in cells that expressed decreased TG2 levels. TG2 knockdown decreased CREB phosphorylation, and CREB knockdown decreased MMP-2 expression. The effect of TG2 on CREB activity and MMP-2 transcription is mediated by TG2-dependent degradation of protein phosphatase 2 (PP2A-alpha). We show that PP2A-alpha complexes with and is targeted for degradation by TG2. In addition to their related in vitro expression levels, TG2 and MMP-2 expression were significantly correlated in vivo, as shown by concordant immunostaining in peritoneal xenografts and in human ovarian tumors. The capacity of TG2 to regulate MMP-2 expression in vitro and in vivo identifies a mechanism that may facilitate tissue invasion and the spread of EOC. The demonstration that TG2 induced degradation of PP2A-alpha activates CREB, and thereby increases MMP-2 transcription, provides novel mechanistic insight into the pro- metastatic function of TG2.

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    • "On the cytoplasmic side of the plasma membrane, TG2 activates phospholipase C (PLC) and RhoA-ROCK-2 signaling pathways [21,22,31], while on the cell surface, TG2 acts as a co-receptor for integrins to promote cell attachments [49]. TG2 is known to positively regulate the expression and activity of metalloproteinases (MMPs)-2 and −9 in cancer cells [10,50]. MMPs are a family of secreted proteins known to promote cancer cell migration through acting in extracellular environment [51]. "
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    ABSTRACT: Tissue-transglutaminase (TG2), a dual function G-protein, plays key roles in cell differentiation and migration. In our previous studies we reported the mechanism of TG2-induced cell differentiation. In present study, we explored the mechanism of how TG2 may be involved in cell migration. To study the mechanism of TG2-mediated cell migration, we used neuroblastoma cells (SH-SY5Y) which do not express TG2, neuroblastoma cells expressing exogenous TG2 (SHYTG2), and pancreatic cancer cells which express high levels of endogenous TG2. Resveratrol, a natural compound previously shown to inhibit neuroblastoma and pancreatic cancer in the animal models, was utilized to investigate the role of TG2 in cancer cell migration. Immunofluorescence assays were employed to detect expression and intracellular localization of TG2, and calcium levels in the migrating cells. Native gel electrophoresis was performed to analyze resveratrol-induced cellular distribution and conformational states of TG2 in migrating cells. Data are presented as the mean and standard deviation of at least 3 independent experiments. Comparisons were made among groups using one-way ANOVA followed by Tukey-Kramer ad hoc test. TG2 containing cells (SHYTG2 and pancreatic cancer cells) exhibit increased cell migration and invasion in collagen-coated and matrigel-coated transwell plate assays, respectively. Resveratrol (1 muM-10 muM) prevented migration of TG2-expressing cells. During the course of migration, resveratrol increased the immunoreactivity of TG2 without affecting the total TG2 protein level in migrating cells. In these cells, resveratrol increased calcium levels, and depletion of intracellular calcium by a calcium chelator, BAPTA, attenuated resveratrol-enhanced TG2 immunoreactivity. In native-polyacrylamide gels, we detected an additional TG2 protein band with slower migration in total cell lysates of resveratrol treated cells. This TG2 form is non-phosphorylated, exclusively present in plasma membrane fractions and sensitive to intracellular Ca2+ concentration suggesting a calcium requirement in TG2-regulated cell migration. Taken together, we conclude that resveratrol induces conformational changes in TG2, and that Ca2+-mediated TG2 association with the plasma membrane is responsible for the inhibitory effects of resveratrol on cell migration.
    Full-text · Article · Apr 2014 · BMC Cancer
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    • "Previous studies have indicated that the AKT/SP-1 pathway regulated MMP-2 promoter activity and affected the migration ability of cancer cells [24,37]. Satpathy et al showed that tissue transglutaminase 2 modulates CREB activation and MMP-2 transcription in ovarian cancer [38]. The upstream promoter sequence of the MMP-9 gene contains AP-1 and NF-κB sites. "
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    ABSTRACT: Crude extracts of Selaginellatamariscina, an oriental medicinal herb, have been evidenced to treat several human diseases. This study investigated the mechanisms by which Selaginellatamariscina inhibits the invasiveness of human oral squamous-cell carcinoma (OSCC) HSC-3 cells. Herein, we demonstrate that Selaginellatamariscina attenuated HSC-3 cell migration and invasion in a dose-dependent manner. The anti-metastatic activities of Selaginellatamariscina occurred at least partially because of the down-regulation of matrix metalloproteinases (MMP)-2 and MMP-9 gelatinase activity and the down-regulation of protein expression. The expression and function of both MMP-2 and MMP-9 were regulated by Selaginellatamariscina at a transcriptional level, as shown by quantitative real-time PCR and reporter assays. Chromatin immunoprecipitation (ChIP) data further indicated that binding of the cAMP response element-binding (CREB) protein and activating protein-1 (AP-1) to the MMP-2 promoter diminished at the highest dosage level of Selaginellatamariscina. The DNA-binding activity of specificity protein 1 (SP-1) to the MMP-9 promoter was also suppressed at the same concentration. Selaginellatamariscina did not affect the mitogen-activated protein kinase signaling pathway, but did inhibit the effects of gelatinase by reducing the activation of serine-threonine kinase Akt. These results demonstrate that Selaginellatamariscina may be a potent adjuvant therapeutic agent in the prevention of oral cancer.
    Full-text · Article · Jun 2013 · PLoS ONE
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    • "Treatment with rTG2 induced formation of networks, indicative of an invasive phenotype (Figure 4A, lower panels). SKOV3 cells that express TG2 and stably transfected with vector or AS-TG2 [8] [14] were used as a second model. Control cells formed networks, whereas AS-TG2 cells remained clumped. "
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    ABSTRACT: Tissue transglutaminase (TG2) is a multifunctional protein that binds to fibronectin and exerts protein transamidating activity in the presence of Ca(2+). We previously reported that TG2 is upregulated in ovarian tumors and enhances intraperitoneal (i.p.) metastasis. TG2 is secreted abundantly in ovarian cancer (OC) ascites as an active enzyme, yet its function in the extracellular compartment remains unknown. To study the distinct functions of secreted TG2, we used recombinant His6-tagged TG2 and catalytically inactive enzyme in vitro and in vivo. By using i.p. and orthotopic ovarian xenografts, we show that extracellular transglutaminase promoted OC peritoneal metastasis. The main pathway activated by extracellular TG2 was noncanonical nuclear factor-kappa B (NF-κB) signaling, and the enzymatic function of the protein was required to induce phosphorylation of IκB kinase α and processing of the precursor protein p100 into the active p52 subunit. A specific target of TG2-activated p52/RelB complex is the hyaluronan receptor, CD44. Noncanonical NF-κB activation by extracellular TG2 induced CD44 up-regulation and epithelial-to-mesenchymal transition, contributing to increased cancer cell invasiveness and OC peritoneal dissemination. Taken together, our data support that noncanonical NF-κB activation is the pathway through which extracellular TG2 promotes OC metastasis.
    Preview · Article · Jun 2013 · Neoplasia (New York, N.Y.)
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