目的:我们和其他人先前已经表明细胞融合在癌症转移中起重要作用。用光谱不同的荧光蛋白对癌症和基质细胞进行颜色编码是一种强大的工具,由我们的实验室率先检测细胞融合。我们先前已经报道了通过使用颜色编码的EL4鼠淋巴瘤细胞和表达光谱不同的荧光蛋白的宿主小鼠在转移部位的癌细胞和基质细胞之间的颜色编码细胞融合。细胞融合发生在癌细胞之间或,在癌细胞和正常细胞之间,比如巨噬细胞,成纤维细胞,和间充质干细胞。在本研究中,目的是对在表达绿色荧光蛋白(GFP)的转基因小鼠中由表达红色荧光蛋白(RFP)的细胞形成的原发性肿瘤和多个转移瘤EL4中观察到的融合杂交细胞进行形态学分类,在同基因模型中。
方法:体外培养表达RFP的EL4小鼠淋巴瘤细胞。收集EL4-RFP细胞,并腹膜内注射到具有免疫能力的转基因C57/BL6-GFP小鼠中,以建立同系模型。两周后,处死小鼠,收获每个器官,培养,并使用共聚焦显微镜观察。
结果:EL4腹膜内肿瘤(原发性)和肺转移,肝脏,血,形成了骨髓。收获并培养所有肿瘤。在所有标本中,RFP-EL4细胞,GFP基质细胞,并观察到融合的黄色荧光杂交细胞。融合的杂交细胞表现出各种形态。在肝转移和循环血液中,免疫细胞样圆形黄色荧光融合细胞有随时间减少的趋势。相反,成纤维细胞样纺锤形黄色荧光融合细胞在腹膜内原发性肿瘤中增加,肺转移瘤,还有骨髓.
结论:EL4-RFP细胞和GFP基质细胞之间的细胞融合发生在原发性肿瘤和所有转移部位。融合的杂合细胞的形态在原代和转移部位有所不同。目前的结果表明,融合的癌症和不同形态的基质杂交细胞可能在癌症进展中起重要作用。
OBJECTIVE: We and others have previously shown that cell fusion plays an important role in cancer metastasis. Color coding of cancer and stromal cells with spectrally-distinct fluorescent proteins is a powerful tool, as pioneered by our laboratory to detect cell fusion. We have previously reported color-coded cell fusion between cancer cells and stromal cells in metastatic sites by using color-coded EL4 murine lymphoma cells and host mice expressing spectrally-distinct fluorescent proteins. Cell fusion occurred between cancer cells or, between cancer cells and normal cells, such as macrophages, fibroblasts, and mesenchymal stem cells. In the present study, the aim was to morphologically classify the fusion-hybrid cells observed in the primary tumor and multiple metastases EL4 formed from cells expressing red fluorescent protein (RFP) in transgenic mice expressing green fluorescent protein (GFP), in a syngeneic model.
METHODS: RFP-expressing EL4 murine lymphoma cells were cultured in vitro. EL4-RFP cells were harvested and injected intraperitoneally into immunocompetent transgenic C57/BL6-GFP mice to establish a syngeneic model. Two weeks later, mice were sacrificed and each organ was harvested, cultured, and observed using confocal microscopy.
RESULTS: EL4 intraperitoneal tumors (primary) and metastases in the lung, liver, blood, and bone marrow were formed. All tumors were harvested and cultured. In all specimens, RFP-EL4 cells, GFP-stromal cells, and fused yellow-fluorescent hybrid cells were observed. The fused hybrid cells showed various morphologies. Immune cell-like round-shaped yellow-fluorescent fused cells had a tendency to decrease with time in liver metastases and circulating blood. In contrast fibroblast-like spindle-shaped yellow-fluorescent fused cells increased in the intraperitoneal primary tumor, lung metastases, and bone marrow.
CONCLUSIONS: Cell fusion between EL4-RFP cells and GFP stromal cells occurred in primary tumors and all metastatic sites. The morphology of the fused hybrid cells varied in the primary and metastatic sites. The present results suggest that fused cancer and stromal hybrid cells of varying morphology may play an important role in cancer progression.