Effects of overexpression of membrane-bound transferrin-like protein (MTf) on chondrogenic differentiation in Vitro.
Membrane-bound transferrin-like protein (MTf) is expressed in parallel with the expression of cartilage-characteristic genes during differentiation of chondrocytes, and the MTf level is much higher in cartilage than in other tissues. To investigate the role of MTf in cartilage, we examined the effects of growth factors on MTf expression in mouse prechondrogenic ATDC5 cells and the effect of MTf overexpression on differentiation of ATDC5 and mouse pluripotent mesenchymal C3H10T1/2 cells. In ATDC5 cultures, bone morphogenetic protein-2 and transforming growth factor-beta as well as insulin induced MTf mRNA expression when these peptides induced chondrogenic differentiation. Forced expression of rabbit MTf in ATDC5 cells induced aggrecan, type II collagen, matrilin-1, type X collagen mRNAs, and cell-shape changes from fibroblastic cells to spherical chondrocytes. Accordingly, the synthesis and accumulation of proteoglycans were higher in MTf-expressing cultures than in control cultures. These effects of MTf overexpression correlated with the MTf protein level on the cell surface and decreased in the presence of anti-MTf antibody. However, the aggrecan mRNA level in the ATDC5 cells overexpressing MTf was lower than that in wild type ATDC5 cells exposed to 10 microg/ml insulin. MTf overexpression in C3H10T1/2 cells also induced aggrecan and/or type II collagen mRNA but not the spherical phenotype. These findings suggest that the expression of MTf on the cell surface facilitates the differentiation of prechondrogenic cells, although MTf overexpression alone seems to be insufficient to commit pluripotent mesenchymal cells to the chondrocyte lineage.
Suardita
Ketut
K
Department of Dental and Medical Biochemistry, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Japan.
Fujimoto
Katsumi
K
Oda
Ryo
R
Shimazu
Atsushi
A
Miyazaki
Kazuko
K
Kawamoto
Takeshi
T
Kato
Yukio
Y
D053674:Aggrecans; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000502:physiology; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D019902:Chondrocytes / Q000166:cytology*; D024043:Collagen Type II / Q000235:genetics; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D005786:Gene Expression Regulation / Q000502:physiology; D066298:In Vitro Techniques; D007328:Insulin / Q000502:physiology; D007334:Insulin-Like Growth Factor I / Q000502:physiology; D037181:Lectins, C-Type; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D011509:Proteoglycans / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D015398:Signal Transduction; D014168:Transferrin / Q000235:genetics* / Q000276:immunology; D016212:Transforming Growth Factor beta / Q000502:physiology
C508320:Acan protein, mouse; D053674:Aggrecans; D000911:Antibodies, Monoclonal; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D024043:Collagen Type II; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D007328:Insulin; D037181:Lectins, C-Type; D008565:Membrane Proteins; D011509:Proteoglycans; D012333:RNA, Messenger; D014168:Transferrin; D016212:Transforming Growth Factor beta; D007334:Insulin-Like Growth Factor I