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MiRNA profiling of HPCX1 linked prostate cancer patients and their healthy brothers

ABSTRACT: MicroRNA expression levels in the lymphoblastic cells of prostate cancer patients and their healthy brothers from HPCX1 linked prostate cancer families were analyzed to trace variants that might alter miRNA expression and explain partly an inherited genetic predisposion to prostate cancer.

ORGANISM(S): Homo sapiens

PROVIDER: GSE24201 | GEO | 2011/12/31

SECONDARY ACCESSION(S): PRJNA133705

REPOSITORIES: GEO

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miRNA profiling of HPCX1 linked prostate cancer patients and their healthy brothers

Project description:MicroRNA expression levels in the lymphoblastic cells of prostate cancer patients and their healthy brothers from HPCX1 linked prostate cancer families were analyzed to trace variants that might alter miRNA expression and explain partly an inherited genetic predisposion to prostate cancer. MicroRNA expression levels in lymphoblastoid cell lines were detected using an Agilent Human miRNA V2 Oligo Microarray Kit (Agilent Technologies).100 ng of total RNA was used as a starting material, and miRNAs were labeled using the Agilent miRNA Labeling Kit. Labeled RNA was hybridized to Agilent miRNA arrays with eight identical arrays per slide, with each array containing probes directed against 723 human and 76 human viral miRNAs. Slides were scanned (Agilent microarray scanner) after hybridization and data was extracted using Feature Extraction software, version 9.5.1. (Agilent Technologies). Altogether 14 cancer patients and 15 healthy brothers from 11 families were included with no replicates.

2011-12-31 | E-GEOD-24201 | biostudies-arrayexpress

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Project description:In this study, nonsense-mediated mRNA decay (NMD) inhibition was used for the discovery of truncating mutations. Six prostate cancer (PRCA) patients and their healthy brothers were selected from a group of HPCX1-linked families. mRNA was isolated from their lymphoblastic cells after pharmacological treatment. Expression analyses were done using Agilent 44K oligoarrays, and selected genes were screened for mutations by sequencing. In order to identify genes containing inactivating mutations in the Xq27-q28 region, an NMD microarray analysis with Agilent 44K Whole Human Genome oligonucleotide microarrays was performed in the families showing the strongest linkage to HPCX1. Five families were chosen with 6 affected and 6 healthy controls. Lymphoblastoid cell lines were derived from whole blood and treated with emetine to block NMD pathway. Total RNA was extracted from harvested cells (treated and untreated) and labeled with Cy5 and Cy3. Labeled samples were hybridized to the Agilent 44K Whole Human Genome Oligonucleotide Microarrays. After hybridization microarrays were scanned and data was extracted using Feature Extraction software. The candidate genes (n=17) for subsequent sequence analysis were selected according to the microarray analysis.

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De novo homozygous deletion of segmental KAL1 and entire STS cause Kallmann syndrome and X-linked ichthyosis in a Chinese family

Project description:Kallmann syndrome is a genetically heterogeneous condition and a treatable form of male infertility. Defects in KAL1 gene have been implicated in Kallmann syndrome, which can be associated with X-linked ichthyosis in contiguous gene syndromes. In order to uncover the genetic cause of two brothers with Kallmann syndrome and X-linked ichthyosis, a custom semiconductor targeted resequencing panel to detect seventeen Kallmann syndrome causal genes and STS gene was designed. Next-generation sequencing was performed using this panel in the two affected brothers and their normal parents. To validate the result, we applied CytoScanâ¢ HD array, quantitative real-time PCR and direct PCR electrophoresis analysis with the participants. The patients received clinical assessment, human chorionic gonadotropin treatment and follow-up for 39 months. The results showed that the two affected siblings have the same de novo deletion at Xp22.3 including exons 9-14 of KAL1 gene and entire STS gene but showed different phenotypes in some respects. The secondary sex characteristics of the patients were greatly improved after treatment. We firstly reported that a de novo homozygous deletion contribute to KS with bilateral cryptorchidism and unilateral renal agenesis or normal kidney development and developed a cost-effective and reliable semiconductor targeted resequencing panel for genetic diagnosis of Kallmann syndrome in routinely obtained samples. One of the two brothers with Kallmann syndrome and X-linked ichthyosis was analyzed for validation the results of the deletion detected by next-generation sequencing.

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