In vivo implantation of human osteosarcoma cells in nude mice induces bones with human-derived osteoblasts and mouse-derived osteocytes.
Two human osteosarcoma cell lines, Hu09 and OST, were suspended in Matrigel (Becton Dickinson Labware, Bedford, Massachusetts) and implanted subcutaneously in the backs of nude mice. To study phenotypic changes of tumor cells and host cells, expression of mRNA for osteopontin (OPN), osteocalcin (OC), and osteonectin (ON) was analyzed by in situ hybridization. Bone tissue was formed in the tumors derived from Hu09 cells. OPN mRNA was transcribed predominantly in osteocyte-like cells within the bone, whereas OC mRNA was transcribed in osteoblast-like cells that surrounded the bone. ON mRNA was detected in both types of cells. The similarity of the expression pattern of OPN, OC, and ON during osteogenesis of Hu09 cells to that of normal skeletal development suggests that the bone formed in Hu09-implanted mice is the same as normal bone tissue. By DNA-DNA in situ hybridization using a human-specific Alu probe and a mouse-specific m-L1 probe, osteoblast-like cells in Hu09 tumorous bone were, however, of human origin, whereas osteocyte-like cells were of mouse origin. In the tumors derived from OST cells, no osteogenesis was observed during the experimental period, and the expression of OPN, OC, and ON was not detected in tumor cells. An endochondral bone formation was not evident when these cells were simply implanted into muscle tissue. An endochondral bone was, however, reactively induced in the host mUscle tissue either when 1 alpha-hydroxyvitamin D3 and all-transretinoic acid were administered to OST-implanted mice or when Hu09 cells were pretreated with dexamethasone before implantation. Hu09 implantation seems to be a useful tool not only for the study of the differentiation of osteosarcoma cells but also for the investigation of the mechanism of bone formation. This system, using Hu09 and OST, may provide us with a new tool for the isolation of the unidentified factors that induce or inhibit osteogenesis in vivo.
Hara
A
A
Department of Orthopaedic Surgery, Juntendo University, School of Medicine, Tokyo, Japan.
Ikeda
T
T
Nomura
S
S
Yagita
H
H
Okumura
K
K
Yamauchi
Y
Y
D000818:Animals; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000235:genetics; D002448:Cell Adhesion; D017690:Cell Transplantation; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D006801:Humans; D006887:Hydroxycholecalciferols / Q000494:pharmacology; D017403:In Situ Hybridization; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation / Q000473:pathology; D010006:Osteoblasts / Q000166:cytology; D015675:Osteocalcin / Q000235:genetics; D010011:Osteocytes / Q000166:cytology; D010012:Osteogenesis / Q000187:drug effects; D015676:Osteonectin / Q000235:genetics; D053495:Osteopontin; D012516:Osteosarcoma / Q000473:pathology*; D010750:Phosphoproteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D012795:Sialoglycoproteins / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000235:genetics; D014183:Transplantation, Heterologous; D014212:Tretinoin / Q000494:pharmacology
C525718:BMP2 protein, human; C525809:BMP4 protein, human; C525719:Bmp2 protein, mouse; C525810:Bmp4 protein, mouse; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D016207:Cytokines; D006887:Hydroxycholecalciferols; D015676:Osteonectin; D010750:Phosphoproteins; D012333:RNA, Messenger; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D016212:Transforming Growth Factor beta; D015675:Osteocalcin; D053495:Osteopontin; D014212:Tretinoin; D003907:Dexamethasone