Unknown,Transcriptomics,Genomics,Proteomics

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Novel Rank-Based Statistical Methods Reveal MicroRNAs with Differential Expression in Multiple Cancer Types

ABSTRACT: The data collected in this study consisted of 32 paired samples of cancerous and normal origins from 14 different patients and 8 different cancer types. Each pair of samples was taken from different parts of the same tissue in the same patient - one collected and extracted from the tumor and one collected and extracted from adjacent normal tissue. One pair from each of the breast, lymphoma and prostate tissues, 2 pairs from liver and lung tissues and 3 pairs from colon, ovary and testes tissues. 2 of the ovary and testes pairs were technical replicates. We use commercial adjacent tissue tumor-normal total RNA samples purchased from Ambion/ABI. The samples were hybridized to Agilent's miRNA microarray version 2.0. Keywords: miRNA For each microarray analysis, 100ng total RNAs were labeled with Cy3 per Agilent miRNA Micorarray Systems Protocol v1.5. The labeled RNA samples were hybridized onto Agilent miRNA microarray containing 799 miRNAs(Agilent Human miRNA Microarray kit, V2) for 21 hours at 55C. The arrays were then washed at room temperature and scanned to produce the hybridization signals (Agilent miRNA Micorarray Systems Protocol v1.5). The arrays were scanned with extended dynamic range at 5 and 100% PMT using the Agilent scanner (model G2565AA Technical replicate: Sample 7, Sample 29 Technical replicate: Sample 8, Sample 30 Technical replicate: Sample 19, Sample 31 Technical replicate: Sample 20, Sample 32

ORGANISM(S): Homo sapiens

SUBMITTER: Israel Steinfeld

PROVIDER: E-GEOD-14985 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Novel rank-based statistical methods reveal microRNAs with differential expression in multiple cancer types.

Navon Roy R Wang Hui H Steinfeld Israel I Tsalenko Anya A Ben-Dor Amir A Yakhini Zohar Z

PloS one 20091125 11

<h4>Background</h4>MicroRNAs (miRNAs) regulate target genes at the post-transcriptional level and play important roles in cancer pathogenesis and development. Variation amongst individuals is a significant confounding factor in miRNA (or other) expression studies. The true character of biologically or clinically meaningful differential expression can be obscured by inter-patient variation. In this study we aim to identify miRNAs with consistent differential expression in multiple tumor types usi ...[more]

PMID: 19946373

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Project description:A summary of the work associated to these microarrays is the following: MicroRNAs (miRNAs) are small non-coding RNAs involved in RNA silencing that play a role in many biological processes. They are involved in the development of many diseases, including cancer. Extensive experimental data show that they play a role in the pathogenesis of cancer as well as the development of drug resistance during treatment. MiRNA microarrays of sensitive and MTX-resistant HT29 colon cancer cells were performed. The results were analyzed using the GeneSpring GX11.5 software. Differentially expressed microRNAs in resistant cells were identified and miR-224, which was greatly underexpressed and displayed robust raw signal values, was selected for further studies. Putative targets were predicted using TargetScan 5.1 software and intersected with the data from expression microarrays previously performed. This approach allowed us to identify miR-224 targets that were differentially expressed more than 2-fold in resistant cells. Among them, ARL3, CDS2, DCP2, HSPC159, MYST3 and SLC4A4 were validated at the mRNA level by qRT-PCR. Functional assays using an anti-miR against miR-224 desensitized the cells toward MTX, mimicking the resistant phenotype. On the other hand, siRNA treatment against SLC4A4 or incubation of Poly Purine Reverse Hoogsteen (PPRH) hairpins against CDS2 or HSPC159 increased sensitivity to MTX. These results revealed a role for miR-224 and its targets in MTX resistance in HT29 colon cancer cells. KEYWORDS Methotrexate, miRNAs, drug resistance, DHFR Two cell lines are compared in the study, which are HT29 colon cancer cells sensitive to methotrexate and HT29 cells resistant to 10e-5M MTX. Six samples are provided which correspond to 3 samples for the control condition and 3 samples for the resistant condition. Data files from miRNA and mRNA (previously submitted to GEO as GSE11440) microarrays were analyzed with GeneSpring GX11.5 software (Agilent Technologies) to find differentially expressed miRNAs and their cellular target genes in the resistant cell lines compared to their sensitive counterparts.

Project description:Schmitz2014 - RNA triplex formation The model is parameterized using the parameters for gene CCDC3 from Supplementary Table S1. The two miRNAs which form the triplex together with CCDC3 are miR-551b and miR-138. This model is described in the article: Cooperative gene regulation by microRNA pairs and their identification using a computational workflow. Schmitz U, Lai X, Winter F, Wolkenhauer O, Vera J, Gupta SK. Nucleic Acids Res. 2014 Jul; 42(12): 7539-7552 Abstract: MicroRNAs (miRNAs) are an integral part of gene regulation at the post-transcriptional level. Recently, it has been shown that pairs of miRNAs can repress the translation of a target mRNA in a cooperative manner, which leads to an enhanced effectiveness and specificity in target repression. However, it remains unclear which miRNA pairs can synergize and which genes are target of cooperative miRNA regulation. In this paper, we present a computational workflow for the prediction and analysis of cooperating miRNAs and their mutual target genes, which we refer to as RNA triplexes. The workflow integrates methods of miRNA target prediction; triplex structure analysis; molecular dynamics simulations and mathematical modeling for a reliable prediction of functional RNA triplexes and target repression efficiency. In a case study we analyzed the human genome and identified several thousand targets of cooperative gene regulation. Our results suggest that miRNA cooperativity is a frequent mechanism for an enhanced target repression by pairs of miRNAs facilitating distinctive and fine-tuned target gene expression patterns. Human RNA triplexes predicted and characterized in this study are organized in a web resource at www.sbi.uni-rostock.de/triplexrna/. This model is hosted on BioModels Database and identified by: BIOMD0000000530. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

Dataset Information

Novel Rank-Based Statistical Methods Reveal MicroRNAs with Differential Expression in Multiple Cancer Types

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Novel rank-based statistical methods reveal microRNAs with differential expression in multiple cancer types.

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