Gene expression during osteoclast-like cell formation induced by antifusion regulatory protein-1/CD98/4F2 monoclonal antibodies (MAbs): c-src is selectively induced by anti-FRP-1 MAb.
Human blood monocytes can differentiate into osteoclast-like cells when they are cultured in the presence of anti-FRP-1. Messenger (mRNA) expression of markers related to osteoclasts was analyzed during differentiation of osteoclasts from monocytes. As markers related to osteoclasts, we selected cathepsin-K, carbonic anhydrase (CA) II, vacuolar H(+)-ATPase (v-ATPase), vitronectin receptor (VNR), tartrate-resistant acid phosphatase (TRAP), osteopontin (OPN), galectin-3, c-src, c-fos, and c-fms. The mRNAs other than c-src mRNA were expressed in freshly isolated monocytes or monocytes incubated with control antibody or anti-FRP-1 monoclonal antibody (MAb) for 14 days. Of these mRNAs, cathepsin-K, CA II, v-ATPase, VNR, TRAP, OPN, and c-fms mRNAs were expressed at higher levels in the osteoclast-like cells than those in monocytes cultured with control antibody. On the other hand, galectin-3 mRNA was expressed at lower levels in the osteoclast-like cells, and there was no significant difference in c-fos mRNA expression between the monocytes cultured with control antibody and anti-FRP-1 MAb. c-src mRNA could not be detected in monocytes freshly isolated or incubated with control antibody. Surprisingly, expression of c-src mRNA was induced in monocytes by anti-FRP-1 MAb and was detectable as early as 3 h after anti-FRP-1 MAb treatment, indicating that c-src is selectively induced by anti-FRP-1 MAb treatment. Furthermore, the osteoclast-like cells expressed calcitonin receptor. Receptor activator of NF-kappaB (RANK) mRNA was detectable in freshly isolated monocytes or monocytes cultured with control antibody or anti-FRP-1 MAbs. Maximal expression of RANK was observed in osteoclast-like cells. On the other hand, no receptor activator of NF-KB ligand (RANKL) mRNA was detectable in any of the samples, suggesting that anti-FRP-1 mAb can induce osteoclast-like cells from blood monocytes without RANKL.
Higuchi
Y
Y
Department of Microbiology, Mie University School of Medicine, Tsu-Shi, Mie Prefecture, Japan.
Ito
M
M
Tajima
M
M
Higuchi
S
S
Miyamoto
N
N
Nishio
M
M
Kawano
M
M
Kusagawa
S
S
Tsurudome
M
M
Sudo
A
A
Katou
K
K
Uchida
A
A
Ito
Y
Y
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology*; D015415:Biomarkers; D002352:Carrier Proteins / Q000276:immunology*; D002454:Cell Differentiation / Q000502:physiology; D027261:Fusion Regulatory Protein-1; D005786:Gene Expression Regulation / Q000502:physiology*; D016391:Genes, src; D006801:Humans; D007150:Immunohistochemistry; D009000:Monocytes / Q000378:metabolism; D010010:Osteoclasts / Q000378:metabolism*; D010641:Phenotype; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000954:Antigens, Surface; D015415:Biomarkers; D002352:Carrier Proteins; D027261:Fusion Regulatory Protein-1; D012333:RNA, Messenger