Hasegawa Kosuke, Suetsugu Atsushi, Nakamura Miki, Matsumoto Takuro, Kunisada Takahiro, Shimizu Masahito, Saji Shigetoyo, Moriwaki Hisataka, Bouvet Michael, Hoffman Robert M
Gifu University Graduate School of Medicine, Gifu, Japan.
Gifu University Graduate School of Medicine, Gifu, Japan AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California San Diego, San Diego, CA, U.S.A.
Anticancer Res. 2016 Sep;36(9):4443-8. doi: 10.21873/anticanres.10988.
BACKGROUND/AIM: Fluorescence-guided surgery (FGS) of cancer is an emerging technology. We have previously shown the importance of resecting both the tumor and the tumor microenvironment (TME) for curative FGS. We also previously developed a syngeneic model using the mouse lymphoma cell line EL-4, expressing red fluorescent protein (EL-4-RFP), growing in green fluorescent protein (GFP) transgenic mice, which we have used in the present report to develop FGS of the tumor microenvironment.
EL-4-RFP lymphoma cells were injected subcutaneously in C57/BL6 GFP transgenic mice. EL-4-RFP cells subsequently formed tumors by 35 days after cell transplantation. Using the portable hand-held Dino-Lite digital imaging system, subcutaneous tumors were resected by FGS. Resected tumor tissues were visualized with the Olympus FV1000 confocal microscope.
Using the Dino-Lite, subcutaneous tumors and the tumor microenvironment were clearly visualized and resected. In the resected tumor, host stromal cells, including adipocyte-like cells and blood vessels with lymphocytes, were observed by confocal microscopy in addition to cancer cells by color-coded confocal imaging. The cancer cells and stromal cells in the TME were deeply intermingled in a highly-complex pattern.
Color-coded FGS is an effective method to completely resect cancer cells along with the stromal cells in the TME which interact in a highly-complex pattern. Microscopically, cancer cells invade the TME and vice versa. To prevent tumor recurrence, it is necessary to resect the TME along with the tumor.
背景/目的：癌症的荧光引导手术（FGS）是一项新兴技术。我们之前已经表明，切除肿瘤和肿瘤微环境（TME）对于根治性FGS至关重要。我们之前还利用小鼠淋巴瘤细胞系EL-4构建了一种同基因模型，该细胞系表达红色荧光蛋白（EL-4-RFP），并在绿色荧光蛋白（GFP）转基因小鼠体内生长，我们在本报告中利用该模型开展肿瘤微环境的FGS。
将EL-4-RFP淋巴瘤细胞皮下注射到C57/BL6 GFP转基因小鼠体内。细胞移植后35天，EL-4-RFP细胞形成肿瘤。使用便携式手持Dino-Lite数字成像系统，通过FGS切除皮下肿瘤。切除的肿瘤组织用奥林巴斯FV1000共聚焦显微镜观察。
使用Dino-Lite可以清晰地观察并切除皮下肿瘤及其微环境。在切除的肿瘤中，通过共聚焦显微镜观察到，除癌细胞外，宿主基质细胞，包括脂肪样细胞和有淋巴细胞浸润的血管，通过彩色编码共聚焦成像得以显示。TME中的癌细胞和基质细胞以高度复杂的模式深度交织在一起。
彩色编码FGS是一种有效的方法，能够完整切除癌细胞及其在TME中以高度复杂模式相互作用的基质细胞。在显微镜下，癌细胞侵入TME，反之亦然。为防止肿瘤复发，有必要连同肿瘤一起切除TME。
