Quantitative multiplex quantum dot in-situ hybridisation based gene expression profiling in tissue microarrays identifies prognostic genes in acute myeloid leukaemia
| Autor: | Judith A. Hoyland, Eleni Tholouli, John Liu Yin, Sarah A. MacDermott, Richard J. Byers |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
Male Myeloid Microarray Biophysics Biology Biochemistry Young Adult Gene expression Quantum Dots medicine Humans Multiplex Myeloid Ecotropic Viral Integration Site 1 Protein Molecular Biology In Situ Hybridization Aged Homeodomain Proteins Tissue microarray Gene Expression Profiling Cell Biology Middle Aged medicine.disease Prognosis Molecular biology Survival Analysis Neoplasm Proteins Gene expression profiling Leukemia Leukemia Myeloid Acute medicine.anatomical_structure Tissue Array Analysis Female Bone marrow Transcription Factors |
| Zdroj: | Biochemical and biophysical research communications. 425(2) |
| ISSN: | 1090-2104 |
| Popis: | Measurement and validation of microarray gene signatures in routine clinical samples is problematic and a rate limiting step in translational research. In order to facilitate measurement of microarray identified gene signatures in routine clinical tissue a novel method combining quantum dot based oligonucleotide in situ hybridisation (QD-ISH) and post-hybridisation spectral image analysis was used for multiplex in-situ transcript detection in archival bone marrow trephine samples from patients with acute myeloid leukaemia (AML). Tissue-microarrays were prepared into which white cell pellets were spiked as a standard. Tissue microarrays were made using routinely processed bone marrow trephines from 242 patients with AML. QD-ISH was performed for six candidate prognostic genes using triplex QD-ISH for DNMT1, DNMT3A, DNMT3B, and for HOXA4, HOXA9, Meis1. Scrambled oligonucleotides were used to correct for background staining followed by normalisation of expression against the expression values for the white cell pellet standard. Survival analysis demonstrated that low expression of HOXA4 was associated with poorer overall survival (p=0.009), whilst high expression of HOXA9 (p |
| Databáze: | OpenAIRE |
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