RNF187 is Downregulated Following NF-κB Inhibition in Late Erythroblasts.
Beta (β)-thalassaemic erythroblasts grown in vitro have reduced nuclear factor kappa B (NF-κB) pathway gene expression. By inhibiting this pathway in erythroblasts from normal individuals, important downstream genes affected by this inhibition can be identified. Bay 11-7082 is a potent inhibitor of the NF-κB pathway, it acts irreversibly, inhibiting NF-κB activation by blocking tumor necrosis factor alpha (TNF-α)-induced phosphorylation of the inhibitory IκB subunit thereby preventing NF-κB activation. In this study, hematopoietic stem cells were isolated from the peripheral blood of 6 healthy individuals and were then cultured for 14 days in conditions which promote erythroid differentiation. Following erythroid lineage enrichment, these cells were stimulated with TNFα or inhibited with Bay 11-7082. Subsequent RNA isolation and gene expression analyses were performed using pooled cDNA with custom PCR arrays. Genes of interest were examined individually on non-pooled samples. Our data identified RNF187, a RING finger domain gene as being downregulated in response to NF-κB inhibition.
School of Pathology and Laboratory Medicine, University of Western Australia, Nedlands, WA, Australia.|Department of Haematology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Nedlands, WA, Australia.
Luke
L
Forster
School of Pathology and Laboratory Medicine, University of Western Australia, Nedlands, WA, Australia.|Department of Haematology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Nedlands, WA, Australia.
Jill
J
Finlayson
School of Pathology and Laboratory Medicine, University of Western Australia, Nedlands, WA, Australia. Reza.Ghassemifar@health.wa.gov.au.|Department of Haematology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Nedlands, WA, Australia. Reza.Ghassemifar@health.wa.gov.au.|PathWest, Haematology R&D, Locked Bag 2009, Nedlands, WA, 6909, Australia. Reza.Ghassemifar@health.wa.gov.au.
Reza
R
Ghassemifar
D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D015536:Down-Regulation; D004900:Erythroblasts / Q000166:cytology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D009570:Nitriles / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D013450:Sulfones / Q000494:pharmacology*; D015534:Trans-Activators / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*
C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D016328:NF-kappa B; D009570:Nitriles; D013450:Sulfones; D015534:Trans-Activators; D014409:Tumor Necrosis Factor-alpha; C582733:RNF187 protein, human; D044767:Ubiquitin-Protein Ligases
Erythropoiesis; Gene expression; NF-κB; Quantitative real time PCR