scholarly journals Μοριακές προσεγγίσεις μηχανισμών βιοαποδόμησης πολυαρωματικών υδρογονανθράκων

2014 ◽  
Author(s):  
Ελπινίκη Βανδέρα
Keyword(s):  
In Silico ◽  
Joint Genome Institute ◽  
Qrt Pcr

Οι πολυκυκλικοί αρωματικοί υδρογονάνθρακες (PAHs), συγκαταλέγονται στους πιο διαδεδομένους και επίμονους ρυπαντές του περιβάλλοντος με άμεση συνέπεια στην ανθρώπινη υγεία. Για τη βελτιστοποίηση των τεχνικών βιοαποκατάστασης των ρυπασμένων περιοχών, μέσω της εκμετάλλευσης του τεράστιου καταβολικού δυναμικού των μικροοργανισμών, απαραίτητη προϋπόθεση είναι αφενός η αποσαφήνιση των πορειών αποδόμησης των ρυπαντών από χαρακτηρισμένα βακτηριακά στελέχη και αφετέρου η σε βάθος κατανόηση των προσαρμοστικών μηχανισμών που ενεργοποιούνται στα βακτήρια παρουσία των ξενοβιοτικών αυτών ενώσεων. Σκοπός της παρούσης ερευνητικής μελέτης ήταν η αποσαφήνιση της πορείας βιοαποδόμησης του φαινανθρενίου από ένα νέο βακτηριακό στέλεχος, το οποίο αποτελεί νέο είδος του γένους Arthrobacter και έχει ονομαστεί Arthrobacter phenanthrenivorans (Sphe3). Πρόσφατα σε συνεργασία με το Joint Genome Institute (JGI), ολοκληρώθηκε η πλήρης αλληλούχιση του γονιδιώματός του.Μέσω της ανάλυσης in silico του γονιδιώματος, της μελέτης της έκφρασης γονιδίων με την τεχνική qRT-PCR, της υπερέκφρασης πρωτεϊνών, καθώς επίσης και της πρωτεωμικής ανάλυσης με nanoLC/MS-MS σε κύτταρα στελέχους Sphe3 που καλλιεργήθηκαν παρουσία φαινανθρενίου, φθαλικού, γλυκόζης και μίγματος φαινανθρενίου με γλυκόζη, πραγματοποιήθηκε ο χαρακτηρισμός όλων των ενζύμων που εμπλέκονται στην πορεία καταβολισμού του φαινανθρενίου. Πραγματοποιήθηκε ο λειτουργικός χαρακτηρισμός της διοξυγονάσης αρχικής υδροξυλίωσης του φαινανθρενίου, η οποία μάλιστα διαπιστώθηκε οτι εμφανίζει απόλυτη εξειδίκευση ως προς το φαινανθρένιο.Ο λειτουργικός χαρακτηρισμός των δύο γονιδίων της διοξυγονάσης του 1-υδροξυ-2-ναφθοϊκού οξέος ( καταβολικό πλασμίδιο και χρωμόσωμα του στελέχους Sphe3), έδειξε ότι τα ένζυμα που κωδικεύουν ανήκουν στην οικογένεια των cupin πρωτεϊνών και έχουν παρόμοιες καταλυτικές ιδιότητες. Ο περαιτέρω καταβολισμός του 1-υδροξυ-2-ναφθοϊκού οξέος προχωράει είτε μέσω της ortho- είτε μέσω της meta-σχάσης του πρωτοκατεχοϊκού οξέος, όπως φάνηκε από την μελέτη in silico και αποδείχθηκε και από την πρωτεωμική ανάλυση. Η μελέτη των γονιδίων που κωδικεύουν τα ένζυμα της meta-σχάσης σε μεταγραφικό επίπεδο, έδειξε ότι τα γονίδια αυτά συμμεταγράφονται συνιστώντας ένα οργανωμένο οπερόνιο. Πειράματα συν-μεταβολισμού, έδειξαν τη δυνατότητα του στελέχους να συν-καταβολίζει πλήρως το φλουρένιο και το ανθρακένιο και μερικώς το φλουορανθένιο παρουσία του φαινανθρενίου.Τόσο από την πρωτεωμική ανάλυση όσο και από πειράματα qRT-PCR, φαίνεται ότι όλα τα ένζυμα που μετέχουν στην καταβολική πορεία του φαινανθρενίου καταστέλλονται παρουσία της γλυκόζης. Επιπλέον, η πρωτεωμική ανάλυση συνέβαλε και στην κατανόηση των μηχανισμών προσαρμογής του στελέχους Sphe3 παρουσία ξενοβιοτικών ενώσεων. Έτσι, παρουσία των αρωματικών υποστρωμάτων παρατηρήθηκε μεταβολή στα επίπεδα έκφρασης ενζύμων του κεντρικού μεταβολισμού του άνθρακα και του μεταβολισμού των αμινοξέων, πρωτεϊνών μεταφορέων, πρωτεϊνών του κυτταρικού φακέλου και πρωτεϊνών που επάγονται παρουσία καταπόνησης. Η παρούσα πρωτεωμική ανάλυση αποτελεί και την πρώτη μελέτη παρουσία υποστρωμάτων PAHs και γλυκόζης σε στέλεχος Arthrobacter, αφού δεν υπάρχει κάποια πρότερη αναφορά στη διεθνή βιβλιογραφία.

scholarly journals Análisis in silico y expresión génica del factor de transcripción TgNAC01 implicado en xilogénesis y estrés abiótico en Tectona grandis.

2017 ◽  
Vol 22 (3) ◽  
pp. 359-369 ◽  
Author(s):  
Vladimir Camel Paucar ◽  
Esteban Galeano ◽  
Helaine Carrer
Keyword(s):  
In Silico ◽  
Tectona Grandis ◽  
Qrt Pcr ◽  
Expresión Génica ◽  
R Project

El xilema secundario es el componente más abundante de la biomasa vegetal. Por tanto, conocer los genes que regulan su formación ayudaría a diseñar estrategias para el mejoramiento genético de la madera. Así, el objetivo de este trabajo fue realizar el análisis computacional de la estructura primaria y secundaria del factor de transcripción (FT) TgNAC01 de Tectona grandis, además de evaluar su historia evolutiva, dominios conservados y expresión génica en tejidos lignificados de árboles de 12 y 60 años. Para ello, se realizó una evaluación del potencial de interacción ion-electrón (PIIE), mediante el método del espectro de la información (MEI) utilizando la librería SFAPS de R-Project, seguido del modelamiento estructural utilizando el software MODELLER y visualizado mediante PyMol. Además, el análisis de alineamiento de secuencia múltiple y filogenia fue mediante el software Bioedit y MrBayes respectivamente. También se evaluó los niveles de síntesis del FT TgNAC01 mediante qRT-PCR. Como resultados, se evidencio que el FT mantiene una estructura β-hoja antiparalela retorcida, que se compacta contra una α-hélice en la región N-terminal, teniendo así tres dominios α hélice y siete dominios β plegada. Asimismo, mediante el MEI se demostró que tiene alrededor de cinco funciones biológicas y mutaciones sobre los aminoácidos con mayor PIIE, lo que conlleva a evoluciones sobre las redes de regulación genética. Finalmente, el FT TgNAC01 juega un papel fundamental en la organización y desarrollo de las partes que componen la albura, como las células radiales de la zona cambial, los vasos, fibras y los anillos de crecimiento.

scholarly journals Evaluation of WHO listed COVID-19 qPCR primers and probe in silico with 375 SERS-CoV-2 full genome sequences

2020 ◽  
Author(s):  
Derek Toms ◽  
Julang Li ◽  
Hugh Y. Cai
Keyword(s):  
In Silico ◽  
Sequence Data ◽  
World Health ◽  
Sequence Information ◽  
Genome Sequences ◽  
Qrt Pcr ◽  
Reverse Transcription Pcr ◽  
Successful Design ◽  
Pcr Assays ◽  
Health Organization

AbstractQuantitative reverse-transcription PCR (qRT-PCR) assays remains the gold standard for detection of the SARS-CoV-2 virus because of its sensitivity and specificity. However, successful design of qRT-PCR assays requires accurate viral genome sequences. With mutations accumulating as the virus is transmitted globally, we sought to compare current assays recommended by the World Health Organization with available SARS-CoV-2 genomic sequences in silico. While most sequences were conserved, there were notable mismatches, particularly in assays developed using early sequences when compared to more recent isolates. We recommend that any assay being evaluated for diagnostic tests be compared with prevalent sequence data from the region of proposed testing and that continued publicly accessible sequence information continue to be provided by the research community.

scholarly journals Integrated transcriptome, small RNA and degradome sequencing approaches proffer insights into chlorogenic acid (CGA) biosynthesis in leafy sweet potato

2019 ◽  
Author(s):  
Wenying Zhang ◽  
Yi Liu ◽  
Wenjin Su ◽  
Lianjun Wang ◽  
Jian Lei ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Chlorogenic Acid ◽  
Rna Sequencing ◽  
Sweet Potato ◽  
Small Rna ◽  
In Silico ◽  
Small Rna Sequencing ◽  
Degradome Sequencing ◽  
Qrt Pcr ◽  
Dicaffeoylquinic Acid

Abstract Background Phenolic compounds play key roles in health protection and leafy sweet potato is an excellent source of total phenolics (TP). The chlorogenic acid (CGA) family, which includes caffeoylquinic acid (CQA), 3,4-O-dicaffeoylquinic acid (3,4-diCQA), 3,5-O-dicaffeoylquinic acid (3,5-diCQA) and 4,5-O-dicaffeoylquinic acid (4,5-diCQA), constitutes the major components of phenolic compounds in leafy sweet potato. However, the mechanism of CGA biosynthesis in leafy sweet potato is unclear. The objective of present study is to dissect the mechanisms of CGA biosynthesis by using transcriptome, small RNA and degradome sequencing.Results Transcriptome sequencing of twelve samples (three replicates) from one low-CGA content genotype and one high-CGA content genotype at two stages (65 and 85 days after planting) identified a total of 2333 common differentially expressed genes (DEGs). The enriched DEGs were related to photosynthesis, starch and sucrose metabolism and phenylpropanoid biosynthesis. In this study, functional genes CCR , CCoAOMT and HCT in the CGA biosynthetic pathway were uniformly downregulated, indicating the way to lignin was altered. Small RNA sequencing of the samples resulted in the identification of 171 microRNAs (miRNAs), including 149 known and 22 novel miRNAs. A total of 38 miRNAs were differentially expressed. Using in silico approaches, 1799 targets were predicated for 38 DE miRNAs. The target genes were enriched in lignin and phenylpropanoid catabolic process. Transcription factors (TFs) such as APETALA2/ethylene response factor ( AP2/ERF ) and Squamosa promoter binding protein -like ( SPL ) predicated in silico were validated in degradome sequencing. The association analysis of DE miRNAs and transcriptome datasets identified that MIR156 family targeted DHQ / SDH (3-dehydroquinate dehydratase/shikimate dehydrogenase), the key gene in the phenylpropanoid pathway , the result of which was validated by qRT-PCR. The expression trends of genes and miRNAs of qRT-PCR were consistent with the RNA and small RNA sequencing results.Conclusions This study established comprehensive functional genomic resources for the CGA biosynthesis and provided insights into the molecular mechanisms involving in this process. The results also enabled the first insights into the regulatory roles of mRNAs and miRNAs and offered candidate genes for leafy sweet potato improvement.

scholarly journals Computational and in vitro Investigation of miRNA-Gene Regulations in Retinoblastoma Pathogenesis: miRNA Mimics Strategy

2015 ◽  
Vol 9 ◽  
pp. BBI.S21742 ◽  
Author(s):  
Nalini Venkatesan ◽  
Perinkulam Ravi Deepa ◽  
Vikas Khetan ◽  
Subramanian Krishnakumar
Keyword(s):  
Cell Cycle ◽  
In Silico ◽  
Apoptotic Cell ◽  
Minimum Energy ◽  
Mirna Gene ◽  
Stem Cell Markers ◽  
Qrt Pcr ◽  
Mirna Mimic ◽  
In Vitro Investigation

Purpose Retinoblastoma (RB), a primary pediatric intraocular tumor, arises from primitive retinal layers. Several novel molecular strategies are being developed for the clinical management of RB. miRNAs are known to regulate cancer-relevant biological processes. Here, the role of selected miRNAs, namely, miR-532-5p and miR-486-3p, has been analyzed for potential therapeutic targeting in RB. Methods A comprehensive bioinformatic analysis was performed to predict the posttranscriptional regulators (miRNAs) of the select panel of genes [Group 1: oncogenes (HMGA2, MYCN, SYK, FASN); Group 2: cancer stem cell markers (TACSTD, ABCG2, CD133, CD44, CD24) and Group 3: cell cycle regulatory proteins (p53, MDM2)] using Microcosm, DIANALAB, miRBase v 18, and REFSEQ database, and RNA hybrid. The expressions of five miRNAs, namely, miR-146b-5p, miR-532-5p, miR-142-5p, miR-328, and miR-486-3p, were analyzed by qRT-PCR on primary RB tumor samples (n = 30; including 17 invasive RB tumors and 13 noninvasive RB tumors). Detailed complementary alignment between 5’ seed sequence of differentially expressed miRNAs and the sequence of target genes was determined. Based on minimum energy level and piCTAR scores, the gene targets were selected. Functional roles of these miRNA clusters were studied by using mimics in cultured RB (Y79, Weri Rb-1) cells in vitro. The gene targets (SYK and FASN) of the studied miRNAs were confirmed by qRT-PCR and western blot analysis. Cell proliferation and apoptotic studies were performed. Results Nearly 1948 miRNAs were identified in the in silico analysis, From this list, only 9 upregulated miRNAs (miR-146b-5p, miR-305, miR-663b, miR-299, miR-532-5p, miR-892b, miR-501, miR-142-5p, and miR-513b) and 10 downregulated miRNAs (miR-1254, miR-328, miR-133a, miR-1287, miR-1299, miR-375, miR-486-3p, miR-720, miR-98, and miR-122*) were found to be common with the RB serum miRNA profile. Downregulation of five miRNAs (miR-146b-5p, miR-532-5p, miR-142-5p, miR-328, and miR-486-3p) was confirmed experimentally. Predicted common oncogene targets (SYK and FASN) of miR-486-3p and miR-532-5p were evaluated for their mRNA and protein expression in these miRNA mimic-treated RB cells. Experimental overexpression of these miRNAs mediated apoptotic cell death without significantly altering the cell cycle in RB cells. Conclusion Key miRNAs in RB pathogenesis were identified by an in silico approach. Downregulation of miR-486-3p and miR-532-5p in primary retinoblastoma tissues implicates their role in tumorigenesis. Prognostic and therapeutic potential of these miRNA was established by the miRNA mimic strategy.

scholarly journals Sensory Neuropathy Affects Cardiac miRNA Expression Network Targeting IGF-1, SLC2a-12, EIF-4e, and ULK-2 mRNAs

2019 ◽  
Vol 20 (4) ◽  
pp. 991 ◽  
Author(s):  
Péter Bencsik ◽  
Krisztina Kiss ◽  
Bence Ágg ◽  
Júlia Baán ◽  
Gergely Ágoston ◽  
...  
Keyword(s):  
Diastolic Dysfunction ◽  
Mirna Expression ◽  
In Silico ◽  
Glucose Transporter ◽  
Molecular Targets ◽  
Sensory Neuropathy ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Qrt Pcr ◽  
Mrna Targets

Background: Here we examined myocardial microRNA (miRNA) expression profile in a sensory neuropathy model with cardiac diastolic dysfunction and aimed to identify key mRNA molecular targets of the differentially expressed miRNAs that may contribute to cardiac dysfunction. Methods: Male Wistar rats were treated with vehicle or capsaicin for 3 days to induce systemic sensory neuropathy. Seven days later, diastolic dysfunction was detected by echocardiography, and miRNAs were isolated from the whole ventricles. Results: Out of 711 known miRNAs measured by miRNA microarray, the expression of 257 miRNAs was detected in the heart. As compared to vehicle-treated hearts, miR-344b, miR-466b, miR-98, let-7a, miR-1, miR-206, and miR-34b were downregulated, while miR-181a was upregulated as validated also by quantitative real time polymerase chain reaction (qRT-PCR). By an in silico network analysis, we identified common mRNA targets (insulin-like growth factor 1 (IGF-1), solute carrier family 2 facilitated glucose transporter member 12 (SLC2a-12), eukaryotic translation initiation factor 4e (EIF-4e), and Unc-51 like autophagy activating kinase 2 (ULK-2)) targeted by at least three altered miRNAs. Predicted upregulation of these mRNA targets were validated by qRT-PCR. Conclusion: This is the first demonstration that sensory neuropathy affects cardiac miRNA expression network targeting IGF-1, SLC2a-12, EIF-4e, and ULK-2, which may contribute to cardiac diastolic dysfunction. These results further support the need for unbiased omics approach followed by in silico prediction and validation of molecular targets to reveal novel pathomechanisms.

scholarly journals Boosting SARS-CoV-2 qRT-PCR detection combining pool sample strategy and mathematical modeling

2020 ◽  
Author(s):  
Isadora Alonso Correa ◽  
Tamires de Souza Rodrigues ◽  
Alex Queiroz ◽  
Leon de Franca Nascimento ◽  
Thiago Wolff ◽  
...  
Keyword(s):  
Mathematical Modeling ◽  
Statistical Modeling ◽  
In Silico ◽  
Cost Savings ◽  
In Silico Analysis ◽  
Standard Technique ◽  
High Sensitivity ◽  
Molecular Testing ◽  
Testing Program ◽  
Qrt Pcr

qRT-PCR is the gold standard technique available for SARS-CoV-2 detection. However, the long test run time and costs associated with this type of molecular testing are a challenge in the actual pandemic scenario. Due to high testing demand, pooling sample strategy is an interesting approach to allow cost savings. We aim to evaluate pooling tests in experimental procedures, as well as perform in silico statistical modeling analysis validated with specimen samples obtained from a mass testing program of Industry Federation of the State of Rio de Janeiro (Brazil). Although the sensitivity reduction in samples pooled with 32 individuals was observed, the high-test sensitivity is maintained even when 16 and 8 samples were pooled. The in silico analysis showed high-cost savings in populations with positive rates lower than 15.0% according to the pool size. This data was validated with the results obtained in our mass testing program: statistical modeling predicted a cost saving of 48.0%, which in practice, was 51.5%, already considering the expenditures with pool sampling that were analyzed individually. Our data confirmed that mathematical modeling is a powerful strategy to improve the pooling approach for SARS-CoV-2 mass testing around the world while maintaining high sensitivity and robustness.

scholarly journals Caracterización in silico y análisis de la expresión expresión génica de proteínas abundantes en la embriogénesis tardía de Agave tequilana Weber var. azul

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Jorge Villegas-Camas ◽  
Karina Verdel-Aranda ◽  
Joel Lara-Reyna ◽  
Aída Martínez-Hernández
Keyword(s):  
In Silico ◽  
Agave Tequilana ◽  
Qrt Pcr ◽  
Expresión Génica

Conocimiento previo/especie: Agave tequilana Weber var. azul es un importante cultivo en México, utilizado para la producción de tequila. Muchas especies de Agave son tolerantes a condiciones áridas. Sin embargo, las bases moleculares de los mecanismos seleccionados en los agaves para confrontar el estrés abiótico, no han sido descritas. Hipótesis: Las proteínas abundantes en la embriogénesis tardía (LEAPs), una superfamilia asociada a las respuestas ante el estrés abiótico en plantas, son un elemento clave en las respuestas de los agaves ante ambientes áridos. Métodos: Datos transcriptómicos de A. tequilana fueron utilizados para realizar análisis in silico e identificar genes que codifican Agave LEAPs. Comparamos sus características estructurales y su similitud/divergencia con LEAPs de otras plantas, utilizando bioinformática. La abundancia de los transcritos de AteqLEAP en órganos vegetativos y en respuesta a altas temperaturas fue determinada mediante qRT-PCR. Resultados: Identificamos tres AteqLEAPs estructuralmente diferentes. Las AteqLEA_5Bs muestran similitud (relativamente baja) con LEAPs conocidas como “atípicas” (LEA_3) y exhiben, inesperadamente, altos niveles de expresión constitutiva en hojas. Los transcritos de AteqLEA_5C (LEA_2) mostraron baja expresión en todos los órganos analizados. Dos isoformas de AteqDHN tipo SK3 muestran el típico desorden estructural e hidrofilicidad de las dehidrinas y son altamente expresadas en hojas no desarrolladas, meristemo vegetativo y tallo (piña). Conclusiones: Las AteqLEAP_5B parecen tener un papel protector preventivo en las hojas fotosintéticas plenamente funcionales; mientras que las AteqDHNs parecen proteger tejidos en proceso de diferenciación como meristemos y hojas en desarrollo; así como tejidos de almacenamiento, como el tallo del agave.

miR-744/eNOS/NO axis: A novel target to halt triple negative breast cancer progression

2021 ◽  
pp. 1-9
Author(s):  
Farah Hady El Kilany ◽  
Rana Ahmed Youness ◽  
Reem Amr Assal ◽  
Mohamed Zakaria Gad
Keyword(s):  
Breast Cancer ◽  
Cancer Progression ◽  
In Silico ◽  
In Silico Analysis ◽  
No Production ◽  
Cellular Viability ◽  
Griess Reagent ◽  
Normal Tissues ◽  
Qrt Pcr ◽  
Silico Analysis

BACKGROUND: Nitric oxide (NO) may have a dual role in cancer. At low concentrations, endogenous NO promotes tumor growth and proliferation. However, at very high concentrations, it mediates cancer cell apoptosis and inhibits cancer growth. High levels of NO have been observed in blood of breast cancer (BC) patients, which increases tumor blood flow and promotes angiogenesis. To date, the regulation of NO-synthesizing enzyme, eNOS, by miRNAs has not been adequately investigated in BC. Therefore, the main aim of this study is to unravel the possible regulation of eNOS by miRNAs in BC and to examine their influence on NO production and BC progression. METHODS: Expression profile of eNOS in Egyptian BC patients and MDA-MB-231 cell lines was investigated using qRT-PCR. In-silico analysis was performed to predict a putative upstream regulator of eNOS. miR-744-5p was selected and its expression was quantified in BC tissues using qRT-PCR. MDA-MB-231 cells were cultured and transfected with miR-744-5p using lipofection method. NO levels were determined using Griess Reagent. Cellular viability and colony-forming ability were assessed using MTT and colony-forming assays; respectively. RESULTS: eNOS and miR-744-5p were significantly up-regulated in BC tissues compared to paired normal tissues. In-silico analysis revealed that miR-744-5p putatively binds to eNOS transcript with high binding scores. Transfection of MDA-MB-231 cells with miR-744-5p mimics resulted in a significant up-regulation of eNOS and consequently NO levels. In addition, miR-744-5p transfection led to an increase in cellular viability and colony-forming ability of the MDA-MB-231. CONCLUSION: miR-744-5p acts as an upstream positive regulator of the NO synthesizing enzyme, eNOS which in turn elevates NO levels. Furthermore, miR-744-5p is a novel oncogenic miRNA in BC. Thus, targeting miR-744/eNOS/NO axis may act as a therapeutic tool in TNBC.

MicroRNA for circulating tumor cells detection in breast cancer: In silico and in vitro analysis

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e22027-e22027 ◽  
Author(s):  
V. Medina Villaamil ◽  
M. Blanco Calvo ◽  
S. M. Díaz Prado ◽  
S. Antolín Novoa ◽  
L. Calvo Martínez ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Progression ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cell Lines ◽  
In Silico ◽  
Detection Methods ◽  
Computational Tools ◽  
Relative Expression ◽  
Qrt Pcr

e22027 Background: Detection of circulating tumor cells (CTC) may provide diagnostic and prognostic information in breast cancer (bc) patients. Deregulation of microRNAs (miRNAs) is frequent in tumors, including breast cancer. It is suggested miRNAs play a role in tumor progression. We hypothesized that miRNAs may constitute a promising new class of cancer biomarkers for CTC detection. Our objective has been to identify miRNAs potentially useful for CTC detection. Methods: Phase I preclinical study was performed by means of computational tools for miRNAs profiling including MIRGATOR, MIRBASE, SMIRNAdb, GeneHUB-GEPIS, MICRORNA.ORG, and MIRNAMAP. In silico data were used to identify and prioritize miRNAs highly expressed in breast cancer, but absent in hematopoietic-derived sources. Selected miRNAs were evaluated by means of qRT-PCR in breast cancer and Hematopoietic cell lines, normal blood, and blood from breast cancer patients. Results: Computational tools identify a set of miRNAs highly expressed in breast cancer sources in relation to hematopoietic samples. Among these were miR-141, miR-200c, miR-196a, miR-203, miR-200a, miR-200b, miR-32, miR-375, miR-31, miR-193a, and miR-205. For instance, relative expression of miR-32 was 104 higher in bc cell lines (N= 5) than in normal PB (N = 19) using qRT-PCR. In a preliminary analysis of PB from bc patients (stage IV) higher relative expression levels for selected miRNAs were found comparing with age-matched controls’ blood. Conclusions: Our results suggest that miRNA bioinformatic approach is a useful high-throughput method to select bc-associated miRNAS. The selected miRNAs should be further evaluated for their potential as markers for CTC detection. We next investigate if blood miRNA profile could predict tumor progression and survival. Supported by grants PI06/1541 and PI07/0477 from Fondo de Investigaciones Sanitarias (FIS), Instituto de Salud Carlos III. No significant financial relationships to disclose.

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