title: High lung-metastatic variant of human osteosarcoma cells, selected by passage of lung metastasis in nude mice, is associated with increased expression of α(v)β(3) integrin.
abstract: Altered expression of αvβ3 integrin is associated with tumor progression and metastasis in several types of cancer, including metastatic osteosarcoma. In this study, we demonstrate that in vivo passaging of lung metastasis in nude mice can generate an aggressive variant of human osteosarcoma cells. Experimental metastases were established by injecting 143B human osteosarcoma cells, expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm, in the tail vein of nude mice. Lung metastases were harvested under fluorescence microscopy from nude mice to establish cell lines which were then injected via the tail vein of additional nude mice. This procedure was repeated for four passages in order to isolate highly metastatic variant sublines. When the parental and metastatic variants were transplanted orthotopically into the tibia of nude mice, the 143B-LM4 variant had the highest metastatic rate, approximately 18-fold higher than the parent (p<0.01). αvβ3 integrin expression was increased approximately 5.6-fold in 143B-LM4 compared to parental cells (p<0.05). Thus, serial passage of lung metastases created a highly metastatic variant of human osteosarcoma cells which had increased expression of αvβ3 integrin, suggesting that αvβ3 integrin plays an essential role in osteosarcoma metastasis. With this highly metastatic variant overexpressing αvβ3 integrin, it will now be possible to further investigate the mechanism by which αvβ3 integrin facilitates metastasis.
authors.affiliation: AntiCancer. Inc., 7917 Ostrow Street, San Diego, CA 92111, U.S.A. all@anticancer.com.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Tome
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kimura
authors.forename: Hiroki
authors.initials: H
authors.lastname: Maehara
authors.forename: Naotoshi
authors.initials: N
authors.lastname: Sugimoto
authors.forename: Michael
authors.initials: M
authors.lastname: Bouvet
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tsuchiya
authors.forename: Fuminori
authors.initials: F
authors.lastname: Kanaya
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hoffman
mesh_terms: D000818:Animals; D001859:Bone Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000378:metabolism*; D008175:Lung Neoplasms / Q000556:secondary*; D051379:Mice; D008819:Mice, Nude; D012516:Osteosarcoma / Q000473:pathology*
chemical_list: D039302:Integrin alphaVbeta3
keywords: green fluorescent protein; lung metastasis; nude mice; osteosarcoma; overexpression; passage; red fluorescent protein; variants; αvβ3 integrin