Lymph node metastasis of oral cancer visualized in live tissue by green fluorescent protein expression

Shintani, Satoru; Mihara, Mariko; Nakahara, Yuuji; Aida, Tadateru; Tachikawa, Tetsuhiko; Hamakawa, Hiroyuki

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Volume 38, Issue 7 , Pages 664-669, October 2002

Lymph node metastasis of oral cancer visualized in live tissue by green fluorescent protein expression

Satoru Shintani
- Department of Oral and Maxillofacial Surgery, Ehime University School of Medicine, 454 Shitukawa, Shigenobu-chou, Onsen-gun, Ehime, 791-0295 Japan
- Corresponding author. Tel.: +81-89-960-5392; fax: +81-89-960-5396
Mariko Mihara
- Department of Oral and Maxillofacial Surgery, Ehime University School of Medicine, 454 Shitukawa, Shigenobu-chou, Onsen-gun, Ehime, 791-0295 Japan
Yuuji Nakahara
- Department of Oral and Maxillofacial Surgery, Ehime University School of Medicine, 454 Shitukawa, Shigenobu-chou, Onsen-gun, Ehime, 791-0295 Japan
Tadateru Aida
- Department of Oral Pathology, Showa University, School of Dentistry, 2-1-1, Kitasenju, Ohta-Ku, Tokyo, 145-8515, Japan
Tetsuhiko Tachikawa
- Department of Oral Pathology, Showa University, School of Dentistry, 2-1-1, Kitasenju, Ohta-Ku, Tokyo, 145-8515, Japan
Hiroyuki Hamakawa
- Department of Oral and Maxillofacial Surgery, Ehime University School of Medicine, 454 Shitukawa, Shigenobu-chou, Onsen-gun, Ehime, 791-0295 Japan

Received 3 December 2001; accepted 12 December 2001.

Abstract

Here, we report the establishment of a stably transfected cell line which expresses high levels of green fluorescent protein (GFP), thus permitting the detection and visualization of developing tumors and lymph node metastases after injection into nude mice. Cells of the human oral squamous carcinoma cell line (SAS-L1) were transfected with an expression vector containing a cDNA encoding humanized GFP and the neomycin resistance gene. A clone with stable high-level expression of GFP was selected in vitro using G418. To study metastasis formation, GFP-expressing cells were injected orthotopically into the tongue of nude mice. The resultant tumor growth in the tongue and micrometastases in the lymph nodes could be visualized by GFP fluorescence. Therefore a useful model has been developed for the study of oral cancer, firstly to understand the metastatic process and secondly for the evaluation of potential treatments.

Keywords: Oral cancer, Orthotopic injection, Lymph node metastasis, Green fluorescent protein