Abstract
The chemosensitivity of micrometastasis is an important factor in therapeutic approaches to micrometastasis. The protocol in this chapter presents procedures capable of examining the drug sensitivity of micrometastases to anticancer agents, especially those in the peritoneal cavity, lymph nodes, and the lung in mice. The protocol consists of green fluorescent protein (GFP) gene-tagged metastasis models in mice and unique detection devices for GFP. The latter include a small, convenient stereo fluorescent microscope for internal visualization of micrometastases at the cellular level with magnification and a handy GFP detection device for external, noninvasive monitoring of therapeutic effect of a drug without magnification. Mice are injected with GFP-tagged tumor cells and divided into an early and a late administration group according to the timing of drug administration. Early administration starts from 1–2 d postinjection for peritoneal and lung metastases and 2–3 wk after subcutaneous injection for lymph node metastasis, when the micrometastasis macroscopically remains invisible-less than 0.5 mm in maximum diame-ter-as confirmed by the detection device. The results thus obtained indicate that micrometastases are more sensitive to anticancer agents than advanced metastases. This system using GFP-tagged tumor models is an indispensable tool for micrometastasis research.
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UICC (1997) TNM Classification of Malignant Tumors, 5th ed. (Sobin, L. H. and Wittekind, Ch., eds.), Wiley, New York.
Nakanishi, H., Kodera, Y., Yamamura, Y., et al. (2000) Rapid quantitative detection of carcinoembryonic antigen-expressing free tumor cells in the peritoneal cavity of gastric-cancer patients with real-time RT-PCR on the LightCycler. Int. J. Cancer 89, 411–417.
Yoshioka, S., Fujiwara, Y., Sugita, Y., et al. (2002) Real-time rapid reverse transcriptase-polymerase chain reaction for intraoperative diagnosis of lymph node micrometastasis: clinical application for cervical lymph node dissection in esophageal cancers. Surgery 132(1), 34–40.
Ito, S., Nakanishi, H., Hirai, T., et al. (2002) Quantitative detection of CEA expressing free tumor cells in the peripheral blood of colorectal cancer patients during surgery with real-time RT-PCR on a Light Cycler. Cancer Lett. 183(2), 195–203.
Lin, W. C., Pretlow, T. P., Pretlow, T. G. D., and Culp, L. A. (1990) Development of micrometastases: earliest events detected with bacterial LacZ gene-tagged tumor cells. J. Natl. Cancer Inst. 82, 1497–1503.
Kobayashi, K., Nakanishi, H., Masuda, A., Tezuka, N., Mutai., M., and Tatematsu, M. (1997) Sequential observation of micrometastasis formation by bacterial lacZ gene-tagged Lewis lung carcinoma cells. Cancer Lett. 112, 191–198.
Nakanishi, H., Kobayashi, K., Nishimura, T., Inada, K., Tsukamoto, T., and Tatematsu, M. (1999) Chemosensitivity of micrometastases and circulating tumor cells to uracil and tegafur as evaluated using LacZ gene-tagged Lewis lung carcinoma cell. Cancer Lett. 142, 31–41.
Misteli, T. and Spector, D. L. (1997) Applications of the green fluorescent protein in cell biology and biotechnology. Nat. Biotechnol. 15, 961–964.
Chishima, T., Miyagi, Y., Wang, X., et al. (1997) Cancer invasion and micrometastasis visualized in live tissue by green fluorescent protein expression. Cancer Res. 7(10), 2042–2047.
Yang, M., Baranov, E., Jiang, P., et al. (2000) Whole-body optical imaging of green fluorescent protein-expressing tumors and metastases. Proc. Natl. Acad. Sci. USA 97(3), 1206–1211.
Hoffman, R. M. (1999) Orthotopic transplant mouse models with green fluorescent protein-expressing cancer cells to visualize metastasis and angiogenesis. Cancer Metastasis Rev. 17, 271–277.
Hoffman, R. M. (2002) Green fluorescent protein imaging of tumour growth, metastasis, and angiogenesis in mouse models. Lancet Oncol. 3(9), 546–556.
Ito, S., Nakanishi, H., Ikehara, Y., et al. (2001) Real-time observation of micrometastasis formation in the living mouse liver using a green fluorescent protein gene-tagged rat tongue carcinoma cell line. Int. J. Cancer 91, 212–217.
Mochizuki, Y., Nakanishi, H., Kodera, Y., et al. (2004) TNF-α promotes progression of peritoneal metastasis as demonstrated using a green fluorescence protein (GFP)-tagged human gastric cancer cell line. Clin. Exp. Metastasis 21, 39–47.
Kurebayashi, J., Nukatsuka, M., Fujioka, A., et al. (1997) Postsurgical oral administration of uracil and tegafur inhibits progression of micrometastasis of human breast cancer cells in nude mice. Clin. Cancer Res. 3, 653–659.
Nakanishi, H., Abe, A., Inada, K., Tsukamoto, T., Yasui, K., and Tatematsu, M. (1999) Induction of apoptosis in metastatic foci from human gastric cancer xenografts in nude mice and reduction of circulating tumor cells in blood by 5-FU and 1-hexylcarbamoyl-5-fluorouracil. J. Cancer Res. Clin. Oncol. 125, 660–668.
Nakanishi, H., Mochizuki, Y., Kodera, Y., et al. (2003) Chemosensitivity of peritoneal micrometastases as evaluated using a green fluorescence protein (GFP)-tagged human gastric cancer cell line. Cancer Sci. 94(1), 112–118.
Takeuchi, S., Nakanishi, H., Yoshida, K., et al. (2000) Isolation of differentiated squamous and undifferentiated spindle carcinoma cell lines with differing metastatic potential from a 4-nitroquinoline N-oxide-induced tongue carcinoma in a F344 rat. Jpn. J. Cancer Res. 91, 1211–1221.
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Nakanishi, H., Ito, S., Mochizuki, Y., Tatematsu, M. (2005). Evaluation of Chemosensitivity of Micrometastases with Green Fluorescent Protein Gene-Tagged Tumor Models in Mice. In: Blumenthal, R.D. (eds) Chemosensitivity: Volume II. Methods in Molecular Medicine™, vol 111. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-889-7:351
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DOI: https://doi.org/10.1385/1-59259-889-7:351
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-58829-586-6
Online ISBN: 978-1-59259-889-2
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