title: The MSX1 homeobox transcription factor is a downstream target of PHOX2B and activates the Delta-Notch pathway in neuroblastoma.
abstract: Neuroblastoma is an embryonal tumour of the peripheral sympathetic nervous system (SNS). One of the master regulator genes for peripheral SNS differentiation, the homeobox transcription factor PHOX2B, is mutated in familiar and sporadic neuroblastomas. Here we report that inducible expression of PHOX2B in the neuroblastoma cell line SJNB-8 down-regulates MSX1, a homeobox gene important for embryonic neural crest development. Inducible expression of MSX1 in SJNB-8 caused inhibition of both cell proliferation and colony formation in soft agar. Affymetrix micro-array and Northern blot analysis demonstrated that MSX1 strongly up-regulated the Delta-Notch pathway genes DLK1, NOTCH3, and HEY1. In addition, the proneural gene NEUROD1 was down-regulated. Western blot analysis showed that MSX1 induction caused cleavage of the NOTCH3 protein to its activated form, further confirming activation of the Delta-Notch pathway. These experiments describe for the first time regulation of the Delta-Notch pathway by MSX1, and connect these genes to the PHOX2B oncogene, indicative of a role in neuroblastoma biology. Affymetrix micro-array analysis of a neuroblastic tumour series consisting of neuroblastomas and the more benign ganglioneuromas showed that MSX1, NOTCH3 and HEY1 are more highly expressed in ganglioneuromas. This suggests a block in differentiation of these tumours at distinct developmental stages or lineages.
authors.affiliation: Department of Human Genetics, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Revet
authors.forename: Gerda
authors.initials: G
authors.lastname: Huizenga
authors.forename: Alvin
authors.initials: A
authors.lastname: Chan
authors.forename: Jan
authors.initials: J
authors.lastname: Koster
authors.forename: Richard
authors.initials: R
authors.lastname: Volckmann
authors.forename: Peter
authors.initials: P
authors.lastname: van Sluis
authors.forename: Ingrid
authors.initials: I
authors.lastname: Øra
authors.forename: Rogier
authors.initials: R
authors.lastname: Versteeg
authors.forename: Dirk
authors.initials: D
authors.lastname: Geerts
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015536:Down-Regulation; D015972:Gene Expression Regulation, Neoplastic; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D051957:MSX1 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism*; D014410:Tumor Stem Cell Assay
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; D047908:Intracellular Signaling Peptides and Proteins; D051957:MSX1 Transcription Factor; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D000071656:Receptor, Notch3; D051880:Receptors, Notch; D014157:Transcription Factors; C512941:DLK1 protein, human; C485631:HEY1 protein, human; C109220:NBPhox protein; C499374:NOTCH3 protein, human; C064045:delta protein; C093564:Neurogenic differentiation factor 1