RRC ID 42434
Author Okada A, Shiomi T, Aoki Y, Fujiwara M.
Title Phenytoin stimulates chondrogenic differentiation in mouse clonal chondrogenic EC cells, ATDC5.
Journal J Toxicol Sci
Abstract Phenytoin (DPH) is known to affect bone formation. However, the mechanism of this effect has not been well understood. In this study, we evaluated the effects of DPH on cartilage formation in a model system using ATDC5 cells, a clonal murine chondrogenic cell line. Alcian blue staining for cartilage nodules and real-time reverse-transcription polymerase chain reaction for the expression of genes encoding type II collagen, aggrecan, transforming growth factor (TGF)-beta1, bone morphogenetic protein (BMP)-4, parathyroid hormone-related peptide (PTHrP), indian hedgehog (Ihh), and patched (Ptc) were performed in ATDC5 cells cultured with DPH. The ATDC5 cells demonstrated enhanced cartilage formation in cultures with DPH. During promoted chondrogenic differentiation, it was observed that DPH increased the mRNA expression of TGF-beta1, BMP-4, Ihh, and Ptc, in a dose-dependent manner on Days 5 to 15. In contrast, other antiepileptic drugs, phenobarbital and valproic acid had no effects on chondrogenesis in ATDC5 cells and osteogenesis in MC3T3-E1 cells. Our results provide fundamental evidence that DPH has a direct stimulatory effect on cartilage formation by regulating TGF-beta and hedgehog signaling molecules, and that DPH effect on bone formation, including chondrogenesis and osteogenesis, is distinct from other antiepileptic drugs as suggested in clinical settings.
Volume 30(3)
Pages 145-56
Published 2005-8-1
DOI 10.2131/jts.30.145
PII JST.JSTAGE/jts/30.145
PMID 16141649
MeSH 3T3 Cells Aggrecans Animals Anticonvulsants / pharmacology Bone Morphogenetic Protein 4 Bone Morphogenetic Proteins / genetics Calcification, Physiologic / drug effects Cartilage / drug effects Cartilage / metabolism Cell Differentiation / drug effects* Cell Differentiation / genetics Cell Line, Tumor Chondrocytes / drug effects* Chondrocytes / metabolism Chondrogenesis / drug effects* Chondrogenesis / genetics Chondroitin Sulfate Proteoglycans / genetics Clone Cells Collagen Type II / genetics Dose-Response Relationship, Drug Extracellular Matrix Proteins / genetics Gene Expression / drug effects Hedgehog Proteins Lectins, C-Type / genetics Mice Parathyroid Hormone-Related Protein / genetics Patched Receptors Patched-1 Receptor Phenobarbital / pharmacology Phenytoin / pharmacology* Proteins / genetics Proteins / metabolism RNA, Messenger / genetics RNA, Messenger / metabolism Receptors, Cell Surface / genetics Reverse Transcriptase Polymerase Chain Reaction / methods Time Factors Trans-Activators / genetics Valproic Acid / pharmacology
IF 1.737
Human and Animal Cells ATDC5(RCB0565) MC3T3-E1(RCB1126)