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GREEN FLOURESCENT PROTEIN: A NEW LIGHT TO STUDY METASTASIS.

T. Chishima^1,3, M. Yang^1,3,4, Y. Miyagi ², X. Wang⁴ , E. Baranov⁴, Y. Tan⁴, H. Yamaoka ^1,3 , H. Shimada ¹, A. R. Moossa³, and R. M. Hoffman^3,4. ¹Dept of Surg, Yokohama City Univ School of Med, Yokohama ²Dept of Path, Yokohama City Univ School of Med, Yokohama ³Dept of Surg, Univ of Ca, San Diego, CA ⁴AntiCancer, Inc., San Diego, CA

We have established stable high-level green fluorescent protein (GFP)-expressing cell lines in vitro that permit the detection and visualization of distant micrometastases when they are implanted orthotopically in nude mice^1-5. A large series of GFP expressing human cancer lines have been isolated including those of the lung, prostate, breast, colon and melanoma. Stable high-level expression of GFP was initially confirmed in subcutaneously growing tumors. To utilize GFP expression for metastasis studies, fragments of subcutaneously growing tumors, which are comprised of GFP-expressing cells, were implanted by surgical orthotopic implantation (SOI) in nude mice. Subsequent micrometastases were detected in systemic organs and could be visualized by GFP fluorescence in the lung, liver, brain, lymph node, bone and other organs down to the single-cell level. With this fluorescent tool, we detected and visualized for the first time tumor cells at the microscopic level in fresh viable tissue in their host organs even in live animals which enabled us to study physiologically relevant patterns of invasion and micrometastasis. The results with the GFP-transfected tumor cells, combined with the use of SOI, demonstrate a fundamental advance to visualize and study metastasis in process.
1 Chishima, T., et al., Cancer Research 57, 2042-2047, 1997.
2 Chishima, T., et al., Clin & Exp Meta 15, 547-552, 1997.
3 Chishima, T., et al., Anticancer Research 17, 2377-2384, 1997.
4 Chishima, T., et al., Proc. Natl. Acad. Sci. USA 94, 11573-11576, 1997.
5 Chishima, T., et al., In Vitro Cell. Dev. Biol. 33, 745-747, 1997.