[show abstract][hide abstract] 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.
[show abstract][hide abstract] 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
[show abstract][hide abstract] 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
[show abstract][hide abstract] 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
[show abstract][hide abstract] 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.
[show abstract][hide abstract] 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.
[show abstract][hide abstract] 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. · 3.73 Impact Factor
[show abstract][hide abstract] 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.
[show abstract][hide abstract] 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.
[show abstract][hide abstract] 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
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
[show abstract][hide abstract] 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
[show abstract][hide abstract] 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
[show abstract][hide abstract] 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
[show abstract][hide abstract] ABSTRACT: The bromodomain protein BRD4 is involved in cell proliferation and cell cycle progression, primarily through its role in acetylated chromatin-dependent regulation of transcription at targeted loci. Here, we show that BRD4 is frequently downregulated by aberrant promoter hypermethylation in human colon cancer cell lines and primary tumors. Ectopic re-expression of BRD4 in these colon cancer cell lines markedly reduced in vivo tumor growth, suggesting a role of BRD4 in human colon cancer.
Journal of Molecular Medicine 11/2011; 90(5):587-95. · 4.77 Impact Factor
[show abstract][hide abstract] ABSTRACT: KDM6B/JMJD3 is a histone H3 lysine demethylase with an important gene regulatory role in development and physiology. Here, we show that human JMJD3 expression is induced by the active vitamin D metabolite 1α,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) and that JMJD3 modulates the gene regulatory action of this hormone. 1,25(OH)(2)D(3) activates the JMJD3 gene promoter and increases the level of JMJD3 RNA in human cancer cells. JMJD3 upregulation was strictly dependent on vitamin D receptor (VDR) expression and was abolished by cycloheximide. In SW480-ADH colon cancer cells, JMJD3 knockdown or expression of an inactive mutant JMJD3 fragment decreased the induction by 1,25(OH)(2)D(3) of several target genes and of an epithelial adhesive phenotype. Moreover, JMJD3 knockdown upregulated the epithelial-to-mesenchymal transition inducers SNAIL1 and ZEB1 and the mesenchymal markers fibronectin and LEF1, while it downregulated the epithelial proteins E-cadherin, Claudin-1 and Claudin-7. Additionally, JMJD3 knockdown abolished the nuclear export of β-catenin and the inhibition of β-catenin transcriptional activity caused by 1,25(OH)(2)D(3). Importantly, the expression of JMJD3 correlated directly with that of VDR and inversely with that of SNAI1 in a series of 96 human colon tumours. Our results indicate for the first time that an epigenetic gene coding for a histone demethylase such as JMJD3 is a VDR co-target that partially mediates the effects of 1,25(OH)(2)D(3) on human colon.
Human Molecular Genetics 09/2011; 20(23):4655-65. · 7.69 Impact Factor
[show abstract][hide abstract] ABSTRACT: Cumulative data support the role of ΔTAp73 variants in tumorigenic processes such as drug resistance. We evaluate the impact of TP73 isoforms and their putative target genes ABCB1, HMGB1, and CASP1 on the survival of colon cancer patients and the correlation between their expressions.
We determined in 77 colon cancer patients the expression of ΔEx2p73, ΔEx2/3p73, ΔNp73, TAp73, ABCB1, HMGB1, and CASP1 by quantitative real-time reverse transcriptase-PCR. Tumor characteristics, disease-free survival, and overall survival (OS) were examined in each patient. Functional experiments were carried out to check whether ectopic expression of ΔNp73 modifies the proliferation, drug resistance, migration, and invasion properties of colon tumor cells and the expression of ABCB1, HMGB1, and CASP1.
Positive correlations were observed between the expression levels of ΔTAp73 variants and HMGB1. Furthermore, a trend was observed for ABCB1. Overexpression of ΔEx2/3p73 and ΔNp73 isoforms was significantly associated with advanced stages (P = 0.04 and P = 0.03, respectively) and predicted shortened OS (P = 0.04 and P = 0.05, respectively). High levels of ABCB1 and HMGB1 were associated with shorter OS (P = 0.04 and P = 0.05, respectively). Multivariate analysis showed that, in addition to the tumor stage, ABCB1 and HMGB1 had independent relationships with OS (P = 0.008). Ectopic expression of ΔNp73 was associated with an increase in proliferation and drug resistance.
The positive correlation between ΔTAp73 variants and HMGB1 and ABCB1 expression supports them as TP73 targets. The fact that upregulation of ΔTAp73 isoforms was associated with shortened OS, increase in proliferation, and drug resistance confirms their oncogenic role and plausible value as prognostic markers. ABCB1 and HMGB1, putative ΔTAp73 target genes, strongly predict OS in an independent manner, making clear the importance of studying downstream TP73 targets that could predict the outcome of colon cancer patients better than ΔTAp73 variants themselves do.
Clinical Cancer Research 08/2011; 17(18):6029-39. · 7.84 Impact Factor
[show abstract][hide abstract] ABSTRACT: The presence of free nucleic acids in plasma has been detected in cancer patients and is associated with poor prognosis. In the present study, the mRNA levels of three genes (EPAS1, KIAA0101 and UBE2D3) in plasma from colorectal cancer patients were analyzed. These genes were selected from a previous study of genomic profiles, discriminating between healthy controls and colorectal cancer patients. mRNA levels were analyzed by real-time PCR in the plasma of 154 patients with colorectal cancer. The association of plasma mRNA levels with clinicopathological parameters and patient survival were analyzed. High levels of EPAS1 in the plasma were associated with patients aged over 50 years, relapse of disease and patient mortality. When patients were divided into two groups, early (I and II) and advanced (III and IV) stages, an association was observed between high levels of EPAS1 mRNA and worse disease-free and overall survival in advanced stages. The expression of KIAA0101 and UBE2D3 was not associated with poor prognosis. Thus, our results suggest that EPAS1 mRNA levels may be an indicator of poor prognosis in colorectal cancer patients at advanced stages, obtained by a non-invasive method.
[show abstract][hide abstract] ABSTRACT: The identification of tumour biomarkers that detect the presence of disease using noninvasive diagnostic procedures is a key part of cancer research. We determined in plasma the vesicle-related microRNA (miRNA) expression profile of nonsmall cell lung cancer (NSCLC) and evaluate whether plasma miRNAs can be both discriminating (between patients and healthy controls) and prognostic markers. 365 human miRNAs were analysed by Taqman® low-density arrays (Applied Biosystems, Foster City, CA, USA) in the plasma from 28 NSCLC patients and 20 controls. Five selected miRNAs (let-7f, miR-20b, miR-30e-3p, miR-223 and miR-301) were validated independently by real-time PCR in plasma from 78 NSCLC and 48 controls and correlated with pathologic parameters and survival. Levels of let-7f, miR-20b and miR-30e-3p were decreased in plasma vesicles of NSCLC patients. Moreover, levels of let-7f and miR-30e-3p distinguished between two groups of patients for stage of disease and therefore possibility of surgery. Plasma levels of miR-30e-3p and let-7f were associated with short disease-free survival and overall survival, respectively. NSCLC patients and healthy controls differ in vesicle-related miRNAs in plasma. Levels of let-7f and miR-30e-3p in NSCLC patients are associated with poor outcome. Thus, plasma vesicle-related miRNAs obtained by noninvasive methods could serve as circulating tumour biomarkers of discriminating and prognostic value.
European Respiratory Journal 03/2011; 37(3):617-23. · 6.36 Impact Factor