Downregulation of fibronectin transcription in highly metastatic adenocarcinoma cells

Werbajh, Santiago; Urtreger, Alejandro J.; Puricelli, Lydia; Lustig, Eugenia Sacerdote de; Joffé, Elisa Bal de Kier; Kornblihtt, Alberto R.

doi:10.1016/s0014-5793(98)01473-2

Cited by 29 publications

(28 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In support of this, we showed that, under serum-starved conditions, the C3.6 cells were less adherent, consistent with ErbB-2 promoting anchorage-independent growth (Harris et al, 1999). The apparent rescue of this adhesion phenotype by plating onto fibronectin (Figure 3) substantiates the notion that ErbB-2 suppression of FN1 plays a functional role in anchorage-independent growth and metastasis, as reported previously (Werbajh et al, 1998;Ignatoski et al, 2000). While our data also suggested that HRGb1 can modify adhesion, and hence the potential for invasiveness, the mechanistic details of this modification are unclear and require further detailed investigation.…”

Section: Discussionsupporting

confidence: 89%

Cellular responses to ErbB-2 overexpression in human mammary luminal epithelial cells: comparison of mRNA and protein expression

et al. 2004

View full text Add to dashboard Cite

Microarray analysis offers a powerful tool for studying the mechanisms of cellular transformation, although the correlation between mRNA and protein expression is largely unknown. In this study, a microarray analysis was performed to compare transcription in response to overexpression of the ErbB-2 receptor tyrosine kinase in a model mammary luminal epithelial cell system, and in response to the ErbB-specific growth factor heregulin b1. We sought to validate mRNA changes by monitoring changes at the protein level using a parallel proteomics strategy, and report a surprisingly high correlation between transcription and translation for the subset of genes studied. We further characterised the identified targets and relate differential expression to changes in the biological properties of ErbB-2-overexpressing cells. We found differential regulation of several key cell cycle modulators, including cyclin D2, and downregulation of a large number of interferon-inducible genes, consistent with increased proliferation of the ErbB-2-overexpressing cells. Furthermore, differential expression of genes involved in extracellular matrix modelling and cellular adhesion was linked to altered adhesion of these cells. Finally, we provide evidence for enhanced autocrine activation of MAPK signalling and the AP-1 transcription complex. Together, we have identified changes that are likely to drive proliferation and anchorage-independent growth of ErbB-2-overexpressing cancer cells.

show abstract

Section: Discussionsupporting

confidence: 89%

Cellular responses to ErbB-2 overexpression in human mammary luminal epithelial cells: comparison of mRNA and protein expression

et al. 2004

View full text Add to dashboard Cite

show abstract

“…38 Down-regulation of fibronectin transcription has also been shown to be associated with a highly metastatic form of murine mammary adenocarcinoma cells. 39 These authors demonstrated that loss of fibronectin expression in these cells is not due to changes in the fibronectin promoter sequence but is the result of negative transcriptional regulation. 39 More recent studies using cDNA arrays to identify genes associated with the more metastatic variant of MM3 adenocarcinoma cells as compared to the parent tumor (M3) found that fibronectin was one of only two genes significantly down-regulated among the genes analyzed in the study.…”

Section: Discussionmentioning

confidence: 99%

KSHV Regulation of Fibulin-2 in Kaposi's Sarcoma

Alcendor

Knobel

Desai

et al. 2011

The American Journal of Pathology

View full text Add to dashboard Cite

Kaposi's sarcoma is an angioproliferative tumor caused by Kaposi's sarcoma-associated herpesvirus (KSHV) infection of vascular endothelial cells. Fibulins, proteins that associate with extracellular matrix (ECM) proteins, may have both tumor-suppressive and oncogenic activities. We found that the expression of fibulin-2 protein and mRNA were decreased 50-fold and 26-fold, respectively, in 10-day KSHVinfected dermal microvascular endothelial cells (DMVEC). Using quantitative RT-PCR, we found a fivefold and 25-fold decrease of fibulin-2 extracellular matrix binding partners, fibronectin and tropoelastin, respectively. Time-course transcriptional analyses over 10 days showed that in addition to that of fibulin-2, expression of fibulins 3 and 5 was decreased in KSHVinfected DMVEC, fibulins 1C/1D were increased, and fibulins 4, 6, and 7 were unchanged. KSHV latencyassociated nuclear antigen (LANA) transcription levels rose consistently over the same period. Addition of recombinant fibulin-3 or -5 for 48 hours to 10-day KSHV-infected cells caused a suppression of KSHVinduced vascular endothelial growth factor (VEGF) protein and mRNA levels. Recombinant fibulin-3 also significantly reduced VEGF receptor 3 expression. In pleural effusion lymphoma cell lines that express variable levels of KSHV lytic replication, we observed no detectable fibulin-2 or -5 expression. Finally, fibulin-2 expression was decreased in tissue microarrays from KSHV-infected, LANA-positive patient cells as compared to that in patient nontumor controls. Understanding the interactions between KSHV and the fibulins may lead to the development of novel therapies for treatment of Kaposi's sarcoma.

show abstract

“…The set of primers used for FN EDI (22), FN EDI minigene (20), and FN IIICS (23) were described previously. The set of primers used for EDII amplification was hEDB-dir (16) and mEDB-rev (5Ј-CAGT-GGACAGTGAATGAGTTGG-3Ј).…”

Section: Methodsmentioning

confidence: 99%

Mammary Epithelial-Mesenchymal Interaction Regulates Fibronectin Alternative Splicing via Phosphatidylinositol 3-Kinase

Blaustein

Pelisch

Coso

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

The way alternative splicing is regulated within tissues is not understood. A relevant model of this process is provided by fibronectin, an important extracellular matrix protein that plays a key role in cell adhesion and migration and contains three alternatively spliced regions known as EDI, EDII, and IIICS. We used a cell culture system to simulate mammary epithelial-stromal communication, a process that is crucial for patterning and function of the mammary gland, and studied the effects of extracellular signals on the regulation of fibronectin pre-mRNA alternative splicing. We found that soluble factors from a mammary mesenchymal cell-conditioned medium, as well as the growth factors HGF/SF (hepatocyte growth factor/scatter factor), KGF (keratinocyte growth factor), and aFGF (acidic fibroblast growth factor), stimulate EDI and IIICS but not EDII inclusion into fibronectin mRNA in the mammary epithelial cell line SCp2, favoring fibronectin isoforms associated with proliferation, migration, and tissue remodeling. We explored the signaling pathways involved in this regulation and found that the mammary mesenchymal cell-conditioned medium and HGF/SF act through a phosphatidylinositol 3-kinase-dependent cascade to alter fibronectin alternative splicing. This splicing regulation is independent from promoter structure and de novo protein synthesis but does require two exonic elements within EDI. These results shed light on how extracellular stimuli are converted into changes in splicing patterns.Pre-mRNA alternative splicing is a widespread process that regulates gene expression and is the most important source of protein diversity in vertebrates (1). Fibronectin (FN), 1 the best characterized extracellular matrix (ECM) glycoprotein, plays a key role in cell adhesive and migratory behavior related to fundamental processes such as embryogenesis, wound healing, maintenance of tissue integrity, and malignancy. Different FN polypeptides arise through an intricate pattern of alternative splicing in three regions of the single primary transcript, (from 5Ј to 3Ј) extra domain II (EDII), extra domain I (EDI), and type III connecting segment (IIICS) (also called EDB or EIIIB, EDA or EIIIA, and V region, respectively), resulting in up to 12 variants in rodents. FN alternative splicing is modulated in a cell type-, development-, and age-specific manner and therefore constitutes a paradigm for studying the regulation of this complex process (2). EDI and EDII are cassette exons, either excluded from or included into the mature FN mRNA. The third site of alternative splicing, IIICS, is subject to total inclusion, partial inclusion, or total exclusion due to the presence of an internal 3Ј splice site within this exon.In vivo EDIϩ FN is poorly represented in the ECM of adult normal tissues. However, this variant is over-expressed in developing embryos, wound healing, liver fibrosis, ovary granulosa cell proliferation, and some tumors (2, 3). It has been proposed that EDI inclusion may mediate a conformational change in the whole m...

show abstract

Downregulation of fibronectin transcription in highly metastatic adenocarcinoma cells

Cited by 29 publications

References 27 publications

Cellular responses to ErbB-2 overexpression in human mammary luminal epithelial cells: comparison of mRNA and protein expression

Cellular responses to ErbB-2 overexpression in human mammary luminal epithelial cells: comparison of mRNA and protein expression

KSHV Regulation of Fibulin-2 in Kaposi's Sarcoma

Mammary Epithelial-Mesenchymal Interaction Regulates Fibronectin Alternative Splicing via Phosphatidylinositol 3-Kinase

Contact Info

Product

Resources

About