Félix Bonilla

Hospital Universitario Puerta de Hierro-Majadahonda, Madrid, Spain

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Publications (155)846.3 Total impact

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    ABSTRACT: Adipogenesis requires a differentiation program driven by multiple transcription factors, where PPARγ and C/EBPα play a central role. Recent findings indicate that Snail inhibits adipocyte differentiation in 3T3-L1 and murine mesenchymal stem cells (mMSC). An in-depth quantitative SILAC analysis of the nuclear fraction of Snail-induced alterations of 3T3-L1 cells was carried out. In total, 2251 overlapping proteins were simultaneously quantified in forward and reverse experiments. We observed 574 proteins deregulated by Snail1 using a fold-change ≥1.5, with 111 up- and 463 down-regulated proteins, respectively. Among other proteins, multiple transcription factors such as Trip4, OsmR, Nr2f6, Cbx6 and Prrx1 were down-regulated. Results were validated in 3T3-L1 cells and mMSC cells by western blot and quantitative PCR. Knock-down experiments in 3T3-L1 cells demonstrated that only Nr2f6 (and Trip4 at minor extent) was required for adipocyte differentiation. Ectopic expression of Nr2f6 reversed the effects of Snail1, promoted adipogenesis and inhibited the expression of IL-17. To prove that correlation, we observed that IL-17 and TNFα were among the most up-regulated pro-inflammatory cytokines in Snail-transfected 3T3-L1 and mMSC cells. Furthermore, the blocking of IL-17 activity in Snail-transfected cells promoted adipocyte differentiation, reverting Snail inhibition. In summary, Snail inhibits adipogenesis through a down-regulation of Nr2f6, which in turn facilitates the expression of IL-17, an anti-adipogenic cytokine. These results would support a novel and important role for Snail and Nr2f6 in obesity control.
    Molecular &amp Cellular Proteomics 12/2014; · 7.25 Impact Factor
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    ABSTRACT: Crosstalk between tumor and stromal cells in the tumor microenvironment alter its properties in ways that facilitate the invasive behavior of tumor cells. Here we demonstrate that cancer-associated fibroblasts (CAF) increase the stiffness of the extracellular matrix (ECM) and promote anisotropic fiber orientation, two mechanical signals generated through a Snail1/RhoA/alpha SMA-dependent mechanism that sustains oriented tumor cell migration and invasiveness. Snail1-depleted CAF failed to acquire myofibroblastic traits in response to TGF Beta, including RhoA activation, alpha-SMA-positive stress fibers, increased fibronectin fibrillogenesis and production of a stiff ECM with oriented fibers. Snail1 expression in human tumor-derived CAF was associated with an ability to organize the ECM. In co-culture, a relatively smaller number of Snail1-expressing CAF were capable of imposing an anisotropic ECM architecture, compared to non-activated fibroblasts. Pathologically, human breast cancers with Snail1+ CAF tended to exhibit desmoplastic areas with anisotropic fibers, lymph node involvement and poorer outcomes. Snail1 involvement in driving an ordered ECM was further confirmed in wound healing experiments in mice, with Snail1 depletion preventing the anisotropic organization of granulation tissue and delaying wound healing. Overall, our results showed that inhibiting Snail1 function in CAF could prevent tumor-driven ECM reorganization and cancer invasion. Copyright © 2014, American Association for Cancer Research.
    Cancer research. 12/2014;
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    ABSTRACT: Snail1 transcriptional repressor is a major inducer of epithelial-to mesenchymal transition but is very limitedly expressed in adult animals. We have previously demonstrated that Snail1 is required for the maintenance of mesenchymal stem cells (MSCs), preventing their premature differentiation. Now, we show that Snail1 controls the tumorigenic properties of mesenchymal cells. Increased Snail1 expression provides tumorigenic capabilities to fibroblastic cells; on the contrary, Snail1 depletion decreases tumor growth. Genetic depletion of Snail1 in MSCs that are deficient in p53 tumor suppressor downregulates MSC markers and prevents the capability of these cells to originate sarcomas in immunodeficient SCID mice. Notably, an analysis of human sarcomas shows that, contrarily to epithelial tumors, these neoplasms display high Snail1 expression. This is particularly clear for undifferentiated tumors, which are associated with poor outcome. Together, our results indicate a role for Snail1 in the generation of sarcomas.
    Neoplasia (New York, N.Y.) 06/2014; · 5.48 Impact Factor
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    ABSTRACT: Tumor-derived exosomes mediate tumorigenesis by facilitating tumor growth, metastasis, development of drug resistance, and immunosuppression. However, little is known about the exosomes isolated from bronchoalveolar lavage (BAL) in patients with lung neoplasm. Exosomes isolated in plasma and BAL from 30 and 75 patients with tumor and nontumor pathology were quantified by acetylcholinesterase activity and characterized by Western Blot, Electron Microscopy, and Nanoparticle Tracking Analysis. Differences in exosome cargo were analyzed by miRNA quantitative PCR in pooled samples and validated in a second series of patients. More exosomes were detected in plasma than in BAL in both groups (P < 0.001). The most miRNAs evaluated by PCR array were detected in tumor plasma, tumor BAL, and nontumor BAL pools, but only 56% were detected in the nontumor plasma pool. Comparing the top miRNAs with the highest levels detected in each pool, we found close homology only between the BAL samples of the two pathologies. In tumor plasma, we found a higher percentage of miRNAs with increased levels than in tumor BAL or in nontumor plasma. The data reveal differences between BAL and plasma exosome amount and miRNA content. © 2014 Wiley Periodicals, Inc.
    Genes Chromosomes and Cancer 04/2014; · 3.55 Impact Factor
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    ABSTRACT: Many studies in different biological systems have revealed that 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3) modulates signaling pathways triggered at the plasma membrane by agents such as Wnt, transforming growth factor (TGF)-β, epidermal growth factor (EGF), and others. In addition, 1α,25(OH)2D3 may affect gene expression by paracrine mechanisms that involve the regulation of cytokine or growth factor secretion by neighboring cells. Moreover, post-transcriptional and post-translational effects of 1α,25(OH)2D3 add to or overlap with its classical modulation of gene transcription rate. Together, these findings show that vitamin D receptor (VDR) cannot be considered only as a nuclear-acting, ligand-modulated transcription factor that binds to and controls the transcription of target genes. Instead, available data support the view that much of the complex biological activity of 1α,25(OH)2D3 resides in its capacity to interact with membrane-based signaling pathways and to modulate the expression and secretion of paracrine factors. Therefore, we propose that future research in the vitamin D field should focus on the interplay between 1α,25(OH)2D3 and agents that act at the plasma membrane, and on the analysis of intercellular communication. Global analyses such as RNA-Seq, transcriptomic arrays, and genome-wide ChIP are expected to dissect the interactions at the gene and molecular levels.
    Frontiers in Physiology 01/2014; 5:60.
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    ABSTRACT: Snail1 transcriptional repressor is a major inducer of epithelial-to mesenchymal transition but is very limitedly expressed in adult animals. We have previously demonstrated that Snail1 is required for the maintenance of mesenchymal stem cells (MSCs), preventing their premature differentiation. Now, we show that Snail1 controls the tumorigenic properties of mesenchymal cells. Increased Snail1 expression provides tumorigenic capabilities to fibroblastic cells; on the contrary, Snail1 depletion decreases tumor growth. Genetic depletion of Snail1 in MSCs that are deficient in p53 tumor suppressor downregulates MSC markers and prevents the capability of these cells to originate sarcomas in immunodeficient SCID mice. Notably, an analysis of human sarcomas shows that, contrarily to epithelial tumors, these neoplasms display high Snail1 expression. This is particularly clear for undifferentiated tumors, which are associated with poor outcome. Together, our results indicate a role for Snail1 in the generation of sarcomas.
    Neoplasia. 01/2014;
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    ABSTRACT: Snail1 is a transcriptional factor that plays an important role in epithelial-mesenchymal transition and in the acquisition of invasive properties by epithelial cells. In colon tumours, Snail1 expression in the stroma correlates with lower specific survival of cancer patients. However, the role(s) of Snail1 expression in stroma and its association with patients' survival have not been determined. We used human primary Carcinoma Associated Fibroblasts (CAFs) or Normal Fibroblasts (NFs) and fibroblast cell lines to analyze the effects of Snail1 expression on the pro-tumorigenic capabilities in colon cancer cells. Snail1 expression was higher in CAFs than in NFs and, as well as α-SMA, a classic marker of activated CAFs. Moreover, in tumour samples from 50 colon cancer patients, SNAI1 expression was associated with expression of other CAF markers, such as α-SMA and FAP. Interestingly, co-culture of CAFs with colon cells induced a significant increase in epithelial cell migration and proliferation, which was associated with endogenous SNAI1 expression levels. Ectopic manipulation of Snail1 in fibroblasts demonstrated that Snail1 expression controlled migration as well as proliferation of co-cultured colon cancer cells in a paracrine manner. Furthermore, expression of Snail1 in fibroblasts was required for the co-adjuvant effect of these cells on colon cancer cell growth and invasion when co-xenografted in nude mice. Finally, cytokine profile changes, particularly MCP3 expression, in fibroblasts are put forward as mediators of Snail1-derived effects on colon tumour cell migration. In summary, these studies demonstrate that Snail1 is necessary for the pro-tumorogenic effects of fibroblasts on colon cancer cells. © 2013 Wiley Periodicals, Inc.
    International Journal of Cancer 11/2013; · 6.20 Impact Factor
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    ABSTRACT: Tumor-derived exosomes are emerging as local and systemic cell-to-cell mediators of oncogenic information through the horizontal transfer of mRNAs, microRNAs and proteins during tumorigenesis. The exosomal content has been described as biologically active when taken up by the recipient cell. Identifying the specific molecular cargo of exosomes will help to determine their function in specific steps of the tumorigenic process. Here we evaluate whether ΔNp73 is selectively packaged in tumor-derived exosomes, its function in the acceptor cells in vitro and in vivo and its prognosis potential in cancer. ΔNp73 messenger is enriched in tumor-derived exosomes, suggesting its active sorting in these microvesicles. We observed the transmission of this exosome cargo to different cell types and how it confers proliferation potential and chemoresistance to the acceptor cells in vitro and in animal models. Finally, our data support the potential prognostic value of exosomal ΔNp73 in colon cancer patients.
    Human Molecular Genetics 09/2013; · 7.69 Impact Factor
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    ABSTRACT: Cancer-associated fibroblasts (CAFs) actively participate in reciprocal communication with tumor cells and with other cell types in the microenvironment, contributing to a tumor-permissive neighborhood and promoting tumor progression. The aim of this study is the characterization of how CAFs from primary human colon tumors promote migration of colon cancer cells. Primary CAF cultures from 15 primary human colon tumors were established. Their enrichment in CAFs was evaluated by the expression of various epithelial and myofibroblast specific markers. Co-culture assays of primary CAFs with different colon tumor cells were performed to evaluate pro-migratory CAF-derived effects on cancer cells. Gene expression profiles were developed to further investigate CAF characteristics. Co-culture assays showed significant differences in fibroblast-derived paracrine pro-migratory effects on cancer cells. Moreover, association between CAFs' pro-migratory effects on cancer cells and classical fibroblast activation or stemness markers was observed. CAF gene expression profiles were analyzed by microarray to identify deregulated genes in different pro-migratory CAFs. The gene expression signature, derived from the most pro-tumorogenic CAFs, was identified. Interestingly, this "CAF signature" showed a remarkable prognostic value for the clinical outcome of colon cancer patients. Moreover, this prognostic value was validated in an independent series of 142 colon cancer patients, by RT-qPCR, with a set of four genes included in the "CAF signature". In summary, these studies demonstrate for the first time the heterogeneity of primary CAFs' effect on colon cancer cell migration. A CAF gene expression signature able to classify colon cancer patients into high- and low-risk groups was identified.
    Clinical Cancer Research 09/2013; · 7.84 Impact Factor
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    ABSTRACT: Cancer-associated fibroblasts (CAFs) are essential components of the stroma that play a critical role in cancer progression. This study aimed to identify novel CAFs markers that might contribute to the invasion and the prognosis of colorectal cancer. The azoxymethane/dextran sodium sulfate mouse model of sporadic colon cancer represents an adequate source for the isolation of CAFs and normal fibroblasts (NFs). By using the explants technique we purified CAFs and NFs from colon tissues. Whole cell extracts and supernatants were subjected to in-depth quantitative proteomic analysis by tandem mass spectrometry. Further validations of up-regulated proteins in CAFs were carried out by chemokine microarray analysis and immunohistochemistry of mouse and human tissues. Using a fold-change ≥1.4, we found 132 and 125 differentially-expressed proteins in whole cell extracts and supernatants, respectively. We found CAFs-associated pro-inflammatory and desmoplastic signatures. The pro-inflammatory signature was composed of several cytokines. Among them, CCL2 and CCL8 caused an increase in migration and invasion of colorectal cancer KM12 cells. The desmoplastic signature was composed of 30 secreted proteins. In mouse and human samples, expression of LTBP2, CDH11, OLFML3 and, particularly, FSTL1 was significantly increased in the tumoral stroma, without significant expression in the cancer epithelial cells. The combination of CALU and CDH11 stromal expression showed a significant association to disease-free survival and poor prognosis. We have identified LTBP2, CDH11, OLFML3 and FSTL1 as selective biomarkers of cancer stroma and CALU and CDH11 as candidate stromal biomarkers of prognostic significance in colon cancer.
    Clinical Cancer Research 09/2013; · 7.84 Impact Factor
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    ABSTRACT: TP73 is a member of the TP53 family, whose deregulated expression has been reported in a wide variety of cancers and linked to patients' outcome. The fact that TP73 encodes a complex number of isoforms (TAp73 and ΔTAp73) with opposing functions and the cross-talk with other members of the family (TP53 and TP63) make it difficult to determine its clinical relevance. Here, we review the molecular mechanisms driving TAp73 and ΔTAp73 expression and how these variants inhibit or promote carcinogenesis. We also highlight the intricate interplay between TP53 family members. In addition, we comment on current pharmacological approaches targeting the TP73 pathway and those affecting the TAp73/ΔTAp73 ratio. Finally, we discuss the current data available in the literature that provide evidence on the role of TP73 variants in predicting prognosis. To date, most of the studies that evaluate the status levels of TP73 isoforms have been based on limited-size series. Despite this limitation, these publications highlight the correlation between high levels of the oncogenic forms and failure to respond to chemotherapy and/or shorter survival. Finally, we emphasize the need for studies to evaluate the significance of combining the deregulation of various members of the TP53 family in order to define patient outcome or their responsiveness to specific therapies. © 2013 Wiley Periodicals, Inc.
    Genes Chromosomes and Cancer 08/2013; · 3.55 Impact Factor
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    ABSTRACT: Liver metastasis in colorectal cancer is the major cause of cancer related death. To identify and characterize proteins associated with colon cancer metastasis, we have compared the conditioned serum-free medium of highly metastatic KM12SM colorectal cancer cells with the parental, poorly metastatic, KM12C cells using quantitative SILAC analyses on a linear ion trap-Orbitrap Velos mass spectrometer. In total, 1337 proteins were simultaneously identified in SILAC forward and reverse experiments. For quantification, 1098 proteins were selected in both experiments, with 155 proteins showing >1.5 fold-change. About 52% of these proteins were secreted directly or using alternative secretion pathways. GDF15, S100A8/A9 and SERPINI1 showed capacity to discriminate cancer serum samples from healthy controls using ELISA assays. In silico analyses of deregulated proteins in the secretome of metastatic cells showed a major abundance of proteins involved in cell adhesion, migration and invasion. To characterize the tumorigenic and metastatic properties of some top up- and down-regulated proteins we used siRNA silencing and antibody blocking. Knocking-down expression of NEO1, SERPINI1 and PODXL showed a significant effect on cellular adhesion. Silencing or blocking experiments with SOSTDC1, CTSS, EFNA3, CD137L/TNFSF9, ZG16B and MDK caused a significant decrease in migration and invasion of highly metastatic cells. In addition, silencing of SOSTDC1, EFNA3 and CD137L/TNFSF9 reduced liver colonization capacity of KM12SM cells. Finally, the panel of six proteins involved in invasion showed association to poor prognosis and overall survival after dataset analysis of gene alterations. In summary, we have defined a collection of proteins that are relevant to understand the mechanisms underlying adhesion, migration, invasion and metastasis in colorectal cancer.
    Molecular &amp Cellular Proteomics 02/2013; · 7.25 Impact Factor
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    ABSTRACT: Tumor epithelial cells within a tumor coexist with a complex microenvironment in which a variety of interactions between its various components determine the behavior of the primary tumors. Cancer-associated fibroblasts (CAFs) and M2 macrophages, characterized by a high expression of different markers, including α-SMA, FSP1 and FAP, or CD163 and DCSIGN, respectively, are involved in the malignancy of different tumors. In this study, the expression of the above markers in CAF and M2 macrophages was analyzed by RT-PCR and IHC in the normal mucosa and tumor tissue from a cohort of 289 colorectal cancer patients. Expression of CAF markers and of M2 markers is associated with the clinical outcome of colorectal cancer patients. Moreover, the combination of CAF and M2 markers identifies three groups of patients with clear differences in the progression of the disease. This combined variable could be a decisive factor in the survival of advanced-stage patients. Taken together, these analyses demonstrate the prognostic involvement of inter-relationships between DCSIGN, CD163, α-SMA, FSP1 and FAP markers in the survival of colon cancer patients.
    Cancer Science 01/2013; · 3.48 Impact Factor
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    ABSTRACT: Fibroblast growth factor receptor 4 (FGFR4) is vital in early development and tissue repair. FGFR4 expression levels are very restricted in adult tissues, except in several solid tumors including colorectal cancer, which showed overexpression of FGFR4. Here, FGFR4 mutation analysis discarded the presence of activating mutations, other than Arg(388), in different colorectal cancer cell lines and tumoral samples. Stable shRNA FGFR4-silencing in SW480 and SW48 cell lines resulted in a significant decrease in cell proliferation, adhesion, cell migration and invasion. This decrease in the tumorigenic and invasive capabilities of colorectal cancer cells was accompanied by a decrease of Snail, Twist and TGFβ gene expression levels and an increase of E-cadherin, causing a reversion to a more epithelial phenotype, in three different cell lines. In addition, FGFR4-signaling activated the oncogenic SRC, ERK1/2 and AKT pathways in colon cancer cells and promoted an increase in cell survival. The relevance of FGFR4 in tumor growth was supported by two different strategies. Kinase inhibitors abrogated FGFR4-related cell growth and signaling pathways at the same extent than FGFR4-silenced cells. Specific FGFR4-targeting using antibodies provoked a similar reduction in cell growth. Moreover, FGFR4 knock-down cells displayed a reduced capacity for in vivo tumor formation and angiogenesis in nude mice. Collectively, our data support a crucial role for FGFR4 in tumorigenesis, invasion and survival in colorectal cancer. In addition, FGFR4 targeting demonstrated its applicability for colorectal cancer therapy.
    PLoS ONE 01/2013; 8(5):e63695. · 3.53 Impact Factor
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    ABSTRACT: Granulocyte-macrophage colony-stimulating factor (GM-CSF/CSF2) is a cytokine produced in the hematological compartment that may enhance anti-tumor immune responses, mainly by activation of dendritic cells. Here we show that more than a third of human colorectal tumors exhibit aberrant DNA demethylation of the GM-CSF promoter and overexpress the cytokine. Mouse engraftment experiments with autologous and homologous colon tumors engineered to repress the ectopic secretion of GM-CSF revealed the tumor-secreted GM-CSF to have an immune-associated antitumor effect. Unexpectedly, an immune-independent anti-tumor effect was observed that depended on the ectopic expression of GM-CSF receptor subunits by tumors. Cancer cells expressing GM-CSF and its receptor did not develop into tumors when autografted into immunocompetent mice. Similarly, 100% of the patients with human colon tumors that overexpressed GM-CSF and its receptor subunits survived at least 5 years after diagnosis. These data suggest that expression of GM-CSF and its receptor subunits by colon tumors may be a useful marker for prognosis as well as for patient stratification in cancer immunotherapy.
    Cancer Research 10/2012; · 9.28 Impact Factor
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    ABSTRACT: The Snail1 transcriptional repressor plays a key role in triggering epithelial-to-mesenchymal transition. Although Snail1 is widely expressed in early development, in adult animals it is limited to a subset of mesenchymal cells where it has a largely unknown function. Using a mouse model with inducible depletion of Snail1, here we demonstrate that Snail1 is required to maintain mesenchymal stem cells (MSCs). This effect is associated to the responsiveness to transforming growth factor (TGF)-β1 that shows a strong Snail1 dependence. Snail1 depletion in conditional knockout adult animals causes a significant decrease in the number of bone marrow-derived MSCs. In culture, Snail1-deficient MSCs prematurely differentiate to osteoblasts or adipocytes and, in contrast to controls, are resistant to the TGF-β1-induced differentiation block. These results demonstrate a new role for Snail1 in TGF-β response and MSC maintenance.Oncogene advance online publication, 6 August 2012; doi:10.1038/onc.2012.342.
    Oncogene 08/2012; · 8.56 Impact Factor
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    ABSTRACT: BackgroundThe present study was designed to investigate the possible impact of treatment on changes in plasma DNA showing tumor DNA features in breast cancer patients. MethodsThirty-eight patients were included. DNA extracted from tumor and normal tissues, normal blood cells, and plasma was used for molecular studies. Molecular alterations in six polymorphic markers (TH2, D10S197, D16S421, D17S855, D17S654, D9S161), mutations in TP53, and aberrant methylation of the first exon of p16INK4a were used to characterize tumor and plasma DNA. After mastectomy, 26 patients received adjuvant chemotherapy and 12, hormone therapy. ResultsAt least one molecular alteration was detected in 28 tumors, and 9 patients showed more than one concomitant molecular change. Twenty patients (53%) displayed loss of heterozygosity (LOH), 6 (16%) TP53 mutations and 11 (29%) methylation of the first exon of p16INK4a. The same molecular aberration was observed in plasma DNA of 18 patients (14 receiving chemotherapy and 4 hormone therapy). Six months after mastectomy, these molecular alterations persisted in the plasma DNA of 6 patients (5 who had completed adjuvant chemotherapy and 1 under hormone therapy). ConclusionsCirculating plasma DNA present after mastectomy is modified only partially by adjuvant chemotherapy or hormone therapy. FundamentoEste trabajo fue diseñado para estudiar el posible impacto adyuvante sobre las variaciones de ADN libre en plasma con características del ADN tumoral en pacientes con cáncer de marna. MétodosSe estudiaron 38 enfermas. El ADN para el análisis molecular fue extraído de tumor y del correspondiente tejido normal, de linfocitos y del plasma. Se utilizaron como marcadores para caracterizar el ADN 6 microsatélites (TH2, D10S197, D16S421, D17S855, D17S654 y D9S161), las mutaciones en TP53 y metilación aberrante del primer exón de pl6INK4a. Después de la mastectomía, 26 pacientes recibieron quimioterapia adyuvante y 12, hormonoterapia. ResultadosSe detectó al menos una alteración molecular en 28 tumores, y en 9 casos un mismo tumor presentó más de una alteración molecular simultáneamente. Veinte tumores (53%) presentaron pérdidas de heterocigosidad (LOH), 6 tumores (16%) mutaciones en TP53 y 11 tumores (29%), metilación aberrante de p16INK4a. La misma alteración molecular se observó en el ADN plasmático de 18 pacientes, 14 de las cuales recibieron quimioterapia y cuatro, hormonoterapia. Seis meses después de la mastectomía, 6 pacientes mantenían las mismas alteraciones moleculares en el ADN plasmático, cinco correspondientes al grupo de quimioterapia y una al de hormonoterapia. ConclusionesEl ADN libre en plasma, similar al ADN tumoral, puede encontrarse después de la mastectomía y el tratamiento adyuvante sólo modifica parcialmente este parámetro.
    Clinical and Translational Oncology 04/2012; 2(3):141-145. · 1.28 Impact Factor
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    ABSTRACT: A significant proportion of extracellular nucleic acids in plasma circulate highly protected in tumor-specific exosomes, but it is unclear how the release of exosomes is modulated in carcinogenesis. We quantified by cytometry exosomes in plasma of 91 colorectal cancer patients to evaluate their potential as a tumor indicator and their repercussions on diagnosis and prognosis. We examined the involvement of TSAP6, a TP53-regulated gene involved in the regulation of vesicular secretion, in levels of circulating exosomes in plasma of colorectal patients and in HCT116 TP53-(wild-type and null) human colorectal cancer cell lines. The fraction of exosomes in cancer patients was statistically higher than in healthy controls (mean rank ¼ 53.93 vs. 24.35). High levels of exosomes in plasma of patients correlated with high levels of carcino-embryonic antigen (P ¼ 0.029) and with poorly differentiated tumors (P ¼ 0.039) and tended to have shorter overall survival than patients with low levels (P ¼ 0.056). Release of exosomes did not correlate with TSAP6 expression; and regulation of TSAP6 by TP53 was not shown either in tumor samples or in HCT116 cell lines. Although it was not suggested that the TP53/TSAP6 pathway regulates the release of exosomes into the plasma of colorectal cancer patients, the level of circulating exosomes may be used as a tumor indicator, because it correlates with poor prognosis parameters and shorter survival.
    Genes Chromosomes and Cancer 04/2012; 51(4):409-18. · 3.55 Impact Factor
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    ABSTRACT: Humoral response in cancer patients appears early in cancer progression and can be used for diagnosis, including early detection. By using human recombinant protein and T7 phage microarrays displaying colorectal cancer (CRC)-specific peptides, we previously selected 6 phages and 6 human recombinant proteins as tumor-associated antigens (TAAs) with high diagnostic value. After completing validation in biological samples, TAAs were classified according to their correlation, redundancy in reactivity patterns and multiplex diagnostic capabilities. For predictor model optimization, TAAs were reanalyzed with a new set of samples. A combination of three phages displaying peptides homologous to GRN, NHSL1 and SREBF2 and four proteins PIM1, MAPKAPK3, FGFR4 and ACVR2B, achieved an area under the curve (AUC) of 94%, with a sensitivity of 89.1% and specificity of 90.0%, to correctly predict the presence of cancer. For early colorectal cancer stages, the AUC was 90%, with a sensitivity of 88.2% and specificity of 82.6%. In summary, we have defined an optimized predictor panel, combining TAAs from different sources, with highly improved accuracy and diagnostic value for colorectal cancer. This article is part of a Special Issue entitled: Translational Proteomics.
    Journal of proteomics 03/2012; 75(15):4647-55. · 5.07 Impact Factor
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    ABSTRACT: Vitamin D deficiency is associated with the high risk of colon cancer and a variety of other diseases. The active vitamin D metabolite 1α,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) regulates gene transcription via its nuclear receptor (VDR), and posttranscriptional regulatory mechanisms of gene expression have also been proposed. We have identified microRNA-22 (miR-22) and several other miRNA species as 1,25(OH)(2)D(3) targets in human colon cancer cells. Remarkably, miR-22 is induced by 1,25(OH)(2)D(3) in a time-, dose- and VDR-dependent manner. In SW480-ADH and HCT116 cells, miR-22 loss-of-function by transfection of a miR-22 inhibitor suppresses the antiproliferative effect of 1,25(OH)(2)D(3). Additionally, miR-22 inhibition increases cell migration per se and decreases the antimigratory effect of 1,25(OH)(2)D(3) in both cell types. In silico analysis shows a significant overlap between genes suppressed by 1,25(OH)(2)D(3) and miR-22 putative target genes. Consistently, miR-22 inhibition abrogates the 1,25(OH)(2)D(3)-mediated suppression of NELL2, OGN, HNRPH1, RERE and NFAT5 genes. In 39 out of 50 (78%) human colon cancer patients, miR-22 expression was found lower in the tumour than in the matched normal tissue and correlated directly with that of VDR. Our results indicate that miR-22 is induced by 1,25(OH)(2)D(3) in human colon cancer cells and it may contribute to its antitumour action against this neoplasia.
    Human Molecular Genetics 03/2012; 21(10):2157-65. · 7.69 Impact Factor

Publication Stats

4k Citations
846.30 Total Impact Points

Institutions

  • 1988–2014
    • Hospital Universitario Puerta de Hierro-Majadahonda
      Madrid, Spain
  • 2012–2013
    • Centro de Investigaciones Biológicas
      Madrid, Madrid, Spain
  • 2005–2012
    • Spanish National Research Council
      • • Instituto de Investigaciones Biomédicas "Alberto Sols"
      • • Biological Research Centre
      Madrid, Madrid, Spain
  • 1985–2011
    • Universidad Autónoma de Madrid
      • Departamento de Medicina
      Madrid, Madrid, Spain
  • 2004–2009
    • Institute for Biomedical Research “Alberto Sols“
      • Department of Cancer Biology
      Madrid, Madrid, Spain
  • 2003
    • Cornell University
      • Cell and Developmental Biology
      Ithaca, NY, United States
  • 1997–2000
    • Hospital Universitario Santa Cristina
      Madrid, Madrid, Spain