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Imaging the interaction of α v integrin‐GFP in osteosarcoma cells with RFP‐expressing host stromal cells and tumor‐scaffold collagen in the primary and metastatic tumor microenvironment
Author(s) -
Tome Yasunori,
Kiyuna Tasuku,
Uehara Fuminari,
Bouvet Michael,
Tsuchiya Hiroyuki,
Kanaya Fuminori,
Hoffman Robert M.
Publication year - 2019
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.27353
Subject(s) - stromal cell , green fluorescent protein , integrin , transfection , osteosarcoma , stroma , chemistry , cancer research , pathology , biology , microbiology and biotechnology , immunology , cell culture , cell , medicine , immunohistochemistry , biochemistry , genetics , gene
Human osteosarcoma 143B cells were previously stably transfected with an α v integrin green flourescent protein (GFP) vector. 143B cells expressing α v integrin‐GFP were transplanted orthotopically in the tibia of transgenic nude mice ubiquitously expressing red fluorescent protein (RFP). The primary tumors acquired RFP‐expressing stroma and were passaged orthotopically in the tibia in noncolored nude mice, which maintained the RFP stroma. The interaction of α v integrin‐GFP expression in 143B cells with RFP‐expressing host stromal cells was observed by confocal microscopy using the Olympus FV1000. Collagen fibers were imaged simultaneously in reflectance mode. The RFP‐expressing stroma included cancer‐associated fibroblasts (CAFs) and tumor‐associated macrophages (TAMs) which persisted even 3 weeks after passage to nontransgenic nude mice. CAFs expressing RFP were aligned between collagen fibers and cancer cells expressing α v integrin‐GFP. Six weeks after transplantation, pulmonary metastases expressing α v integrin‐GFP could be identified. TAMs expressing RFP accompanied metastasized osteosarcoma cells expressing α v integrin‐GFP in the lung. The current study demonstrates the importance of α v integrin interaction with stromal elements in osteosarcoma.