title: Use of αv Integrin Linked to Green Fluorescent Protein in Osteosarcoma Cells and Confocal Microscopy to Image Molecular Dynamics During Lung Metastasis in Nude Mice.
abstract: BACKGROUND
  We report here imaging of the behavior of αv integrin linked to green fluorescent protein (GFP) in human osteosarcoma cells colonizing the lung of nude mice.
  
  
  METHODS
  143B osteosarcoma cells expressing αv integrin-GFP were generated by transfection of an αv integrin-GFP fusion-gene vector pCMV-AC- ITGAV-GFP. In order to generate experimental lung metastases, 143B osteosarcoma cells (1×10(6)), stably expressing αv integrin-GFP, were injected intravenously via the tail vein. The osteosarcoma cells were transplanted orthotopically in the tibia of nude mice in order to generate spontaneous metastases. Lungs were harvested and imaged by confocal microscopy within 1 hour.
  
  
  RESULTS
  In the experimental lung-metastasis model, extravasating and deformed osteosarcoma cells expressing αv integrin-GFP were observed. Pseudopodia of the osteosarcoma cells contained small puncta of αv integrin-GFP. In early-stage spontaneous lung metastasis, tumor emboli were observed in pulmonary vessels. At high magnification, small αv integrin-GFP puncta were observed in the tumor embolus. In late-stage spontaneous metastasis, tumor emboli were also observed in pulmonary vessels. Invading cancer cells with strong expression of αv integrin-GFP were observed at the margin of the tumor emboli.
  
  
  CONCLUSIONS
  The results of this study demonstrate that molecular dynamics of αv integrin-GFP can be imaged in lung metastasis, which will allow further understanding of the role of αv integrin in this process. The results also suggest a general concept for imaging molecular behavior in vivo.
authors.affiliation: AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A. Department of Orthopedic Surgery, Graduate School of Medicine, University of the Ryukyus, Nishihara, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Tome
authors.affiliation: AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A.
authors.forename: Shuya
authors.initials: S
authors.lastname: Yano
authors.affiliation: Department of Physiology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan.
authors.forename: Naotoshi
authors.initials: N
authors.lastname: Sugimoto
authors.affiliation: AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A.
authors.forename: Sumiyuki
authors.initials: S
authors.lastname: Mii
authors.affiliation: AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A. Department of Orthopedic Surgery, Graduate School of Medicine, University of the Ryukyus, Nishihara, Japan.
authors.forename: Fuminari
authors.initials: F
authors.lastname: Uehara
authors.affiliation: AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A. Department of Orthopaedic Surgery, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Miwa
authors.affiliation: Department of Surgery, University of California, San Diego, CA, U.S.A.
authors.forename: Michael
authors.initials: M
authors.lastname: Bouvet
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tsuchiya
authors.affiliation: Department of Orthopedic Surgery, Graduate School of Medicine, University of the Ryukyus, Nishihara, Japan.
authors.forename: Fuminori
authors.initials: F
authors.lastname: Kanaya
authors.affiliation: AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A. all@anticancer.com.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hoffman
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D049452:Green Fluorescent Proteins / Q000302:isolation & purification / Q000627:therapeutic use; D006801:Humans; D039564:Integrin alphaV / Q000235:genetics* / Q000302:isolation & purification; D008175:Lung Neoplasms / Q000000981:diagnostic imaging* / Q000473:pathology* / Q000556:secondary; D051379:Mice; D018613:Microscopy, Confocal; D056004:Molecular Dynamics Simulation; D009362:Neoplasm Metastasis; D012516:Osteosarcoma / Q000175:diagnosis / Q000000981:diagnostic imaging* / Q000235:genetics / Q000473:pathology; D023041:Xenograft Model Antitumor Assays
chemical_list: D039564:Integrin alphaV; D049452:Green Fluorescent Proteins
keywords: confocal microscopy; green fluorescent protein; lung metastasis; nude mice; osteosarcoma; real-time imaging; αv Integrin