title: Runx2 deficiency in chondrocytes causes adipogenic changes in vitro.
abstract: Runx2 (runt-related transcription factor 2) is an important transcription factor for chondrocyte differentiation as well as for osteoblast differentiation. To investigate the function of Runx2 in chondrocytes, we isolated chondrocytes from the rib cartilage of Runx2-deficient (Runx2-/-) mice and examined the effect of Runx2 deficiency on chondrocyte function and behavior in culture for up to 12 days. At the beginning of the culture, Runx2-/- chondrocytes actively proliferated, had a polygonal shape and expressed type II collagen; these are all characteristics of chondrocytes. However, they gradually accumulated lipid droplets that stained with oil red O and resembled adipocytes. Northern blot analysis revealed that the expression of adipocyte-related differentiation marker genes including PPAR gamma (peroxisome proliferator-activated receptor gamma), aP2 and Glut4 increased over time in culture, whereas expression of type II collagen decreased. Furthermore, the expression of Pref-1, an important inhibitory gene of adipogenesis, was remarkably decreased. Adenoviral introduction of Runx2 or treatment with transforming growth factor-beta, retinoic acid, interleukin-1 beta, basic fibroblast growth factor, platelet-derived growth factor or parathyroid hormone inhibited the adipogenic changes in Runx2-/- chondrocytes. Runx2 and transforming growth factor-beta synergistically upregulated interleukin-11 expression, and the addition of interleukin-11 to the culture medium reduced adipogenesis in Runx2-/- chondrocytes. These findings indicate that depletion of Runx2 resulted in the loss of the differentiated phenotype in chondrocytes and induced adipogenic differentiation in vitro, and show that Runx2 plays important roles in maintaining the chondrocyte phenotype and in inhibiting adipogenesis. Our findings suggest that these Runx2-dependent functions are mediated, at least in part, by interleukin-11.
authors.affiliation: Department of Molecular Medicine, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
authors.forename: Hirayuki
authors.initials: H
authors.lastname: Enomoto
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Furuichi
authors.forename: Akira
authors.initials: A
authors.lastname: Zanma
authors.forename: Kei
authors.initials: K
authors.lastname: Yamana
authors.forename: Carolina
authors.initials: C
authors.lastname: Yoshida
authors.forename: Satoru
authors.initials: S
authors.lastname: Sumitani
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Motomi
authors.initials: M
authors.lastname: Enomoto-Iwamoto
authors.forename: Masahiro
authors.initials: M
authors.lastname: Iwamoto
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Komori
mesh_terms: D033962:Adaptor Protein Complex 2 / Q000378:metabolism; D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D002135:Calcium-Binding Proteins; D002356:Cartilage / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D048429:Cell Size; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D024043:Collagen Type II / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit; D036341:Intercellular Signaling Peptides and Proteins; D017370:Interleukin-11 / Q000378:metabolism; D050356:Lipid Metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009363:Neoplasm Proteins / Q000172:deficiency* / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D033962:Adaptor Protein Complex 2; D002135:Calcium-Binding Proteins; D024043:Collagen Type II; D050659:Core Binding Factor Alpha 1 Subunit; C512925:Dlk1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D017370:Interleukin-11; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D012097:Repressor Proteins; D014157:Transcription Factors; D016212:Transforming Growth Factor beta