The effects of natural and synthetic retinoids on the differentiation of RCJ C5.18 chondrogenic cells.
RCJ C 5.18 (C 5.18) is a chondrogenic clonal cell line which, under standard culture conditions, develops chondroblastic features including the production of a cartilagenous matrix. Retinoic acid (RA) is known to inhibit the chondrogenic differentiation of C 5.18 cells and this may parallel the teratogenic effects of retinoids in vivo; however, the question as to which of the 3 retinoic acid receptors (RAR alpha, beta, gamma) or the 3 retinoid X receptors (RXR alpha, beta, gamma) mediate this RA-induced inhibition remains unanswered. We tested several retinoids with different receptor binding characteristics. Cartilage formation in C 5.18 cultures was evaluated by counting the number of cartilage nodules formed, and by quantitating the glycosaminoglycan content of the cultures using alcian blue staining. All of the retinoids prevented cartilage formation in a dose-dependent manner. Treatment with the retinoids did not affect cell number, thereby ruling out any toxic effects. RA, which binds to all 3 RARs with similar affinity, produced a 50% inhibition (IC50) of cartilage formation at 4 x 10(-10) M. We also tested Ch55, which also binds to all 3 RARs, but with higher affinity than RA. This compound was approximately 10 times more potent than RA (IC50 2 x 10(-11) M). 9-cis RA, which binds to the 3 RARs with affinities similar to RA and also binds to the 3 RXRs, was less active (IC50 8 x 10(-9) M), suggesting that RXR binding interferes with the inhibitory effect of ligand-activated RARs. 9-cis retinal, for which the binding characteristics are unknown, had the same effect as 9-cis RA.(ABSTRACT TRUNCATED AT 250 WORDS)
MRC Group in Periodontal Physiology, Faculty of Dentistry, University of Toronto, Ontario, Canada.
H P
HP
Von Schroeder
Y
Y
Hashimoto
J N
JN
Heersche
D000818:Animals; D001565:Benzoates / Q000633:toxicity; D002356:Cartilage / Q000166:cytology / Q000187:drug effects* / Q000196:embryology; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D002599:Chalcone / Q000031:analogs & derivatives / Q000633:toxicity; D047188:Chalcones; D004305:Dose-Response Relationship, Drug; D006025:Glycosaminoglycans / Q000096:biosynthesis; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D018168:Receptors, Retinoic Acid / Q000378:metabolism*; D000072482:Retinoic Acid Receptor alpha; D047488:Retinoid X Receptors; D012176:Retinoids / Q000633:toxicity*; D013723:Teratogens / Q000494:pharmacology*; D013764:Tetrahydronaphthalenes / Q000633:toxicity; D014157:Transcription Factors; D014212:Tretinoin / Q000031:analogs & derivatives; D000097768:Retinoic Acid Receptor gamma
D001565:Benzoates; D002599:Chalcone; D047188:Chalcones; D006025:Glycosaminoglycans; D018160:Receptors, Cytoplasmic and Nuclear; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha; D047488:Retinoid X Receptors; D012176:Retinoids; D013723:Teratogens; D013764:Tetrahydronaphthalenes; D014157:Transcription Factors; D014212:Tretinoin; D000097768:Retinoic Acid Receptor gamma; C000607073:Rara protein, rat; C082927:retinoic acid binding protein I, cellular; C082928:retinoic acid binding protein II, cellular; C082925:retinoic acid receptor beta; C061133:tamibarotene; C068073:Am 580; C085268:Re 80; C054189:3,5-di-tert-butylchalcone 4'-carboxylic acid