Identification and testing of a gene expression signature of invasive carcinoma cells within primary mammary tumors

Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Cancer Research (Impact Factor: 9.33). 01/2005; 64(23):8585-94. DOI: 10.1158/0008-5472.CAN-04-1136
Source: PubMed


We subjected cells collected using an in vivo invasion assay to cDNA microarray analysis to identify the gene expression profile of invasive carcinoma cells in primary mammary tumors. Expression of genes involved in cell division, survival, and cell motility were most dramatically changed in invasive cells indicating a population that is neither dividing nor apoptotic but intensely motile. In particular, the genes coding for the minimum motility machine that regulates beta-actin polymerization at the leading edge and, therefore, the motility and chemotaxis of carcinoma cells, were dramatically up-regulated. However, ZBP1, which restricts the localization of beta-actin, the substrate for the minimum motility machine, was down-regulated. This pattern of expression implicated ZBP1 as a suppressor of invasion. Reexpression of ZBP1 in metastatic cells with otherwise low levels of ZBP1 reestablished normal patterns of beta-actin mRNA targeting and suppressed chemotaxis and invasion in primary tumors. ZBP1 reexpression also inhibited metastasis from tumors. These experiments support the involvement in metastasis of the pathways identified in invasive cells, which are regulated by ZBP1.

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Available from: Sumanta Goswami
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    • "It facilitates cell invasion via phosphatidylinositol 3-kinase- dependent local accumulation of actin filaments [32]. Increased ENAH/Mena expression levels correlate with invasiveness of breast and salivary gland tumors[33, 34], and are also seen in colorectal cancer and in polyps with high grade dysplasia [35]. TNS1 (up-regulated 14-fold), which is also known as Tensin 1, has actin cross-linking activity and localizes to focal adhesions. "
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    ABSTRACT: Background Transgelin is an actin-binding protein that promotes motility in normal cells. Although the role of transgelin in cancer is controversial, a number of studies have shown that elevated levels correlate with aggressive tumor behavior, advanced stage, and poor prognosis. Here we sought to determine the role of transgelin more directly by determining whether experimental manipulation of transgelin levels in colorectal cancer (CRC) cells led to changes in metastatic potential in vivo. Methods Isogenic CRC cell lines that differ in transgelin expression were characterized using in vitro assays of growth and invasiveness and a mouse tail vein assay of experimental metastasis. Downstream effects of transgelin overexpression were investigated by gene expression profiling and quantitative PCR. Results Stable overexpression of transgelin in RKO cells, which have low endogenous levels, led to increased invasiveness, growth at low density, and growth in soft agar. Overexpression also led to an increase in the number and size of lung metastases in the mouse tail vein injection model. Similarly, attenuation of transgelin expression in HCT116 cells, which have high endogenous levels, decreased metastases in the same model. Investigation of mRNA expression patterns showed that transgelin overexpression altered the levels of approximately 250 other transcripts, with over-representation of genes that affect function of actin or other cytoskeletal proteins. Changes included increases in HOOK1, SDCCAG8, ENAH/Mena, and TNS1 and decreases in EMB, BCL11B, and PTPRD. Conclusions Increases or decreases in transgelin levels have reciprocal effects on tumor cell behavior, with higher expression promoting metastasis. Chronic overexpression influences steady-state levels of mRNAs for metastasis-related genes.
    Full-text · Article · Dec 2016 · BMC Cancer
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    • "Thus, to facilitate analysis of the mechanism of Mena INV dependent growth factor sensitization, we used human cell lines that ectopically express GFP- Mena, GFP-Mena INV , or GFP alone. Mena was found to be upregulated approximately 10-fold at the RNA level in invasive tumor cells collected from primary xenograft tumors, therefore we engineered our cells to express levels roughly comparable to those seen in invasive cells from primary tumors (Wang et al., 2004) (Fig S1). As expected, Mena INV -expressing cells invaded further into 3D collagen gels (Fig 1A). "
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    ABSTRACT: During breast cancer progression, alternative mRNA splicing produces functionally distinct isoforms of Mena, an actin regulator with roles in cell migration and metastasis. Aggressive tumor cell subpopulations express Mena(INV), which promotes tumor cell invasion by potentiating EGF responses. However, the mechanism by which this occurs remains unknown. Here, we report that Mena associates constitutively with the tyrosine phosphatase PTP1B and mediates a novel negative feedback mechanism that attenuates receptor tyrosine kinase signaling. On EGF stimulation, complexes containing Mena and PTP1B are recruited to the EGFR, causing receptor dephosphorylation and leading to decreased motility responses. Mena also interacts with the 5' inositol phosphatase SHIP2, which is important for the recruitment of the Mena-PTP1B complex to EGFR. When Mena(INV) is expressed, PTP1B recruitment to EGFR is impaired, providing a mechanism for growth factor sensitization to EGF, as well as HGF and IGF, and increased resistance to EGFR and Met inhibitors in signaling and motility assays. In sum, we demonstrate that Mena plays an important role in regulating growth factor-induced signaling. Disruption of this attenuation by Mena(INV) sensitizes tumor cells to low growth factor concentrations, thereby increasing the migration and invasion responses that contribute to aggressive, malignant cell phenotypes. © 2015 by The American Society for Cell Biology.
    Full-text · Article · Sep 2015 · Molecular biology of the cell
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    • "However, there are still promising candidates such as MT1-MMP (Chen et al., 1994; Chen, 1996; Sabeh et al., 2009). In addition, upstream players that induce invadopodia formation such as Mena INV , Arg, IL-6, EGFR, and faciogenital dysplasia protein Fgd1, are known to have increased expression levels in various cancers (Ayala et al., 2009; Clark et al., 2009; Li et al., 2010b; Wang et al., 2004, 2007; Gil-Henn et al., 2013). Thus both upstream regulators and structural components of invadopodia present vital opportunities for diagnosis and therapy. "
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