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AntiCancer, Inc., San Diego, California 92111 [M. Y., S. H., P. J., X. W., Y. T., T. C., R. M. H.]; Department of Surgery, Yokohama City University School of Medicine, Yokohama, Japan [M. Y., S. H., T. C., H. S.]; and Department of Surgery, University of California, San Diego, California 92103-8220 [M. Y., S. H., T. C., A. R. M., R. M. H.]
To understand the skeletal metastatic pattern of non-small cell lung cancer, we developed a stable high-expression green fluorescent protein (GFP) transductant of human lung cancer cell line H460 (H460-GFP). The GFP-expressing lung cancer was visualized to metastasize widely throughout the skeleton when implanted orthotopically in nude mice. H460 was transduced with the pLEIN retroviral expression vector containing the enhanced GFP and the neomycin (G418) resistance gene. A stable high GFP-expressing clone was selected in vitro using 800 µg/ml G418. Stable high-level expression of GFP was maintained in s.c.-growing tumors formed after injecting H460-GFP cells in nude mice. To use H460-GFP for visualization of metastasis, fragments of s.c.-growing H460-GFP tumors were implanted by surgical orthotopic implantation in the left lung of nude mice. Subsequent micrometastases were visualized by GFP fluorescence in the contralateral lung, plural membrane, and widely throughout the skeletal system including the skull, vertebra, femur, tibia, pelvis, and bone marrow of the femur and tibia. The use of GFP-expressing H460 cells transplanted by surgical orthotopic implantation revealed the extensive metastatic potential of lung cancer in particular to widely disseminated sites throughout the skeleton. This new metastatic model can play a critical role in the study of the mechanism of skeletal and other metastasis in lung cancer and in screening of therapeutics that prevent or reverse this process.
1 This study was supported in part by U. S. National Cancer Institute Grant R44 CA53963.
2 To whom requests for reprints should be addressed, at AntiCancer, Inc., San Diego, CA 92111. Phone: (619) 654-2555; Fax: (619) 268-4175; E-mail: all@anticancer.com.
Received 6/19/98. Accepted 8/14/98.
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