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Tumor microRNA expression profiling identifies circulating microRNAs for earlier breast cancer detection

ABSTRACT: Due to their role in tumorigenesis and remarkable stability in body fluids, microRNAs (miRNAs) are emerging as a promising diagnostic tool. The aim of this study was to identify tumor miRNA signatures for the discrimination of breast cancer and the intrinsic molecular subtypes, and the study in plasma of the status of the most significant ones in order to identify potential circulating biomarkers for breast cancer detection. MiRNA expression profiling of 1919 human miRNAs was conducted in 122 FFPE breast tumors (31 luminal A, 33 luminal B, 27 Her2 and 31 triple negative) and 11 normal breast tissues using LNA based miRNA microarrays. Breast tumors were divided into a training (n=61) and a test set (n=61). Both series comprised a similar number of samples from each molecular subtype. Differential expression analysis was performed and microarray classifiers were developed with samples from the training set and validated in samples from the test set. The most relevant miRNAs were validated by quantitative PCR and analyzed in plasma from 36 pretreated patients, 47 postreated patients and 26 healthy individuals. In addition, further validation in 114 pretreated patients and 116 healthy individuals was performed.

ORGANISM(S): Homo sapiens

SUBMITTER: Nerea Matamala

PROVIDER: E-GEOD-58606 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Tumor microRNA expression profiling identifies circulating microRNAs for early breast cancer detection.

Matamala Nerea N Vargas María Teresa MT González-Cámpora Ricardo R Miñambres Rebeca R Arias José Ignacio JI Menéndez Primitiva P Andrés-León Eduardo E Gómez-López Gonzalo G Yanowsky Kira K Calvete-Candenas Julio J Inglada-Pérez Lucía L Inglada-Pérez Lucía L Martínez-Delgado Beatriz B Benítez Javier J

Clinical chemistry 20150608 8

<h4>Background</h4>The identification of novel biomarkers for early breast cancer detection would be a great advance. Because of their role in tumorigenesis and stability in body fluids, microRNAs (miRNAs) are emerging as a promising diagnostic tool. Our aim was to identify miRNAs deregulated in breast tumors and evaluate the potential of circulating miRNAs in breast cancer detection.<h4>Methods</h4>We conducted miRNA expression profiling of 1919 human miRNAs in paraffin-embedded tissue from 122 ...[more]

PMID: 26056355

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Project description:In this study, we compared the expression profiles of miRNAs in blood samples from Impaired Fasting Glucose (IFG) and T2D male patients. Healthy adult males with no past history of T2D (n=158) and with desirable cholesterol and blood pressure profiles were enrolled in this study. They were then classified according to fasting glucose levels to have T2D, IFG or as healthy controls (CTL), for comparison of miRNA expression profiles. Employing miRNA microarray, we identified ‘signature miRNAs’ in peripheral blood samples that distinguished IFG and T2D. Eight selected miRNAs were further validated using stem-loop real-time RT-PCR. miR-144 expression was found to be dysregulated in Type 2 Diabetes, wherein its expression was significantly higher than in healthy controls. Insulin receptor substrate 1 (IRS1) has been predicted to be a potential target of miR-144. Consistent with this observation, IRS1 mRNA and protein levels, verified by quantitative real-time PCR and western blotting respectively, were found to be down-regulated. Dysregulation in expression of microRNAs (miRNAs) in various tissues has been linked to a wide spectrum of diseases, including Type 2 Diabetes mellitus (T2D). In this study, we compared the expression profiles of miRNAs in blood samples from Impaired Fasting Glucose (IFG) and T2D male patients. Healthy adult males with no past history of T2D (n=158) and with desirable cholesterol and blood pressure profiles were enrolled in this study. They were then classified according to fasting glucose levels to have T2D, IFG or as healthy controls (CTL), for comparison of miRNA expression profiles. Employing miRNA microarray, we identified ‘signature miRNAs’ in peripheral blood samples that distinguished IFG and T2D. Eight selected miRNAs were further validated using stem-loop real-time RT-PCR. miR-144 expression was found to be dysregulated in Type 2 Diabetes, wherein its expression was significantly higher than in healthy controls. Insulin receptor substrate 1 (IRS1) has been predicted to be a potential target of miR-144. Consistent with this observation, IRS1 mRNA and protein levels, verified by quantitative real-time PCR and western blotting respectively, were found to be down-regulated. Using luciferase assay, we further demonstrated that miR-144 directly targets IRS1 and showed its effects on protein expression via immunocytochemistry. From this cross-sectional study in humans, we have identified signature miRNAs which could explain the pathogenesis of T2D. Whether miRNAs like miR-144 could be potential therapeutic targets for management of T2D will need to be explored by further mechanistic and functional studies. Total RNA (plus miRNAs) was isolated using a modification of the RiboPure™-Blood kit from Ambion (Austin,TX) according to the manufacturer’s protocol. The concentration of total RNA and integrity were determined by using Nano-Drop ND-1000 Spectrophotometry (NanoDrop Tech, Rockland, Del) and gel electrophoresis respectively.

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Dataset Information

Tumor microRNA expression profiling identifies circulating microRNAs for earlier breast cancer detection

Publications

Tumor microRNA expression profiling identifies circulating microRNAs for early breast cancer detection.

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