British Journal of Haematology · 2000 · 45 citations · 13 references
Life SciencesHematologyMolecular BiologyGene ExpressionGene Expression AnalysisHematopathologyCell-type-specific TranscriptomicsIsoform Expression AnalysisMolecular CytogeneticsPediatric LeukemiasAcute LeukemiasAnimal Disease ModelsBcr-abl1 Fusion GeneQuantitative Polymerase Chain ReactionQuantitative Polymerase Chain Reaction AssaysRelative Abundance Analysis
The Ikaros gene is an essential regulator in development and haematopoiesis. Dysregulated Ikaros gene expression participates in leukaemic processes, as evidenced in animal models, and by analyses of blast-cell populations from leukaemic patients. We used real-time quantitative polymerase chain reaction (PCR) to evaluate the relative abundance of several Ikaros transcript isoforms in a variety of leukaemic-cell samples. Total RNA was isolated from bone-marrow or blood-cell samples collected at diagnosis in children or adult patients, 18 of whom had acute myeloblastic leukaemia (AML), 61 of whom had acute lymphoblastic leukaemia (ALL) and 11 of whom had chronic myeloid leukaemia (CML). The ratio (Ik1 + Ik2)/(Ik1 + Ik2 + Ik4 + Ik7 + Ik8) ranged from 13.5% to 85% and was lower (P < 0. 05) in samples from patients with m-bcr-abl ALL. An alternative splicing resulting in the deletion of 30 nucleotides at the end of exon 6 was observed in leukaemic samples, and in normal thymus and bone marrow. Our results are consistent with previous reports and suggest that the pattern of expression of the different human Ikaros isoforms are not homogeneous among different subsets of leukaemias.
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Real-time quantitative RT–PCR after laser-assisted cell picking
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