title: BMP-6 and retinoic acid synergistically differentiate the IMR-32 human neuroblastoma cells.
abstract: BACKGROUND
  We studied the effects of all-trans retinoic acid (ATRA) and Bone morphogenetic protein-6 (BMP-60 on neuronal differentiation of IMR-32 cells.
  
  
  METHODS
  Immunostaining, Western blotting and MTT assays were used to characterize the survival and neuronal phenotype of IMR-32 cells differentiated by ATRA +/- BMP-6.
  
  
  CONCLUSIONS
  ATRA, BMP-6 and ATRA + BMP-6 treatments each induced dopaminergic neuronal differentiation of the IMR-32 cells as judged by expression of GAP-43, tyrosine hydroxylase and microtubule-associated protein 2ab. ATRA alone induced strong biochemical differentiation, whereas ATRA + BMP-6 treatment synergistically enhanced neurite outgrowth and morphological maturation. When compared with undifferentiated controls, ATRA induced Bcl-2 expression, while loss of Bax expression was observed only in cells differentiated by ATRA + BMP-6. The high Bcl-2/Bax ratio in cells differentiated by ATRA and ATRA + BMP-6, respectively, correlated with the survival of these cells in serum-free media. Upon re-exposure to fresh serum lacking factors, only cells differentiated by ATRA + BMP-6 were unable to resume cell division. In concordance with their morphological maturation, these cells were terminally differentiated when compared with cells differentiated by separate treatment with ATRA or BMP-6.
authors.affiliation: Institute of Anatomy and Cell Biology, Göteborg University, Box 420, SE-405 30 Gothenburg, Sweden.
authors.forename: Venil N
authors.initials: VN
authors.lastname: Sumantran
authors.forename: Anke
authors.initials: A
authors.lastname: Brederlau
authors.forename: Keiko
authors.initials: K
authors.lastname: Funa
mesh_terms: D001685:Biological Factors / Q000494:pharmacology; D015415:Biomarkers; D055418:Bone Morphogenetic Protein 6; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004357:Drug Synergism; D019922:GAP-43 Protein / Q000096:biosynthesis / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D019254:Genes, bcl-2; D016259:Genes, myc; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics; D016501:Neurites / Q000187:drug effects; D009447:Neuroblastoma / Q000473:pathology*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis; D016212:Transforming Growth Factor beta; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis / Q000235:genetics; D051028:bcl-2-Associated X Protein
chemical_list: C494826:BAX protein, human; C525816:BMP6 protein, human; C525819:BMP7 protein, human; D001685:Biological Factors; D015415:Biomarkers; D055418:Bone Morphogenetic Protein 6; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D016895:Culture Media, Serum-Free; D019922:GAP-43 Protein; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D016271:Proto-Oncogene Proteins c-myc; D016212:Transforming Growth Factor beta; D051028:bcl-2-Associated X Protein; D014212:Tretinoin; D014446:Tyrosine 3-Monooxygenase