Investigating the role of macrophages in tumor formation using a MaFIA mouse model
Investigating the role of macrophages in tumor formation using a MaFIA mouse model
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DOI：
10.3892/or.2013.2508
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发表时间：
2013-08-01
期刊：
影响因子：
4.2
通讯作者：
O'Neill, K.
中科院分区：
文献类型：
--
作者：
Clifford, A. B.;Elnaggar, A. M.;O'Neill, K.
肿瘤相关巨噬细胞（TAM）在其发育、生长和转移活动中与肿瘤相互作用。使用允许选择性消耗巨噬细胞的转基因小鼠模型，我们能够获得巨噬细胞促进转移的潜力。在MaFIA（巨噬细胞Fas诱导的细胞凋亡）小鼠中，骨髓谱系的转基因表达细胞在药物AP 20187存在下通过细胞凋亡经历死亡。增强型绿色荧光蛋白（EGFP）融合到自杀基因，以允许转基因表达细胞的鉴定。通过皮下和静脉注射B16-F10黑色素瘤细胞完成肿瘤诱导。将巨噬细胞耗尽的小鼠中的转移与正常对照小鼠中的转移进行比较。检查肺和肾的转移细胞。与对照组相比，巨噬细胞去除组显示出显著较少的转移（P>0.001）。我们推测巨噬细胞可能通过与黑色素瘤细胞融合来帮助转移过程。使用适当的细胞标记物和荧光激活细胞分选，我们能够检测到一小群双阳性细胞。我们通过显微镜分析证实了细胞融合，通过免疫组织化学和免疫荧光观察了细胞的形态。双阳性细胞的存在表明巨噬细胞/癌细胞融合可能是转移的可能机制。
Tumor-associated macrophages (TAMs) interact with tumors in their development, growth and metastatic activities. Using a transgenic mouse model that allows for the selective depletion of macrophages we were able to access the macrophage's potential to facilitate metastasis. In the MaFIA (Macrophage Fas-Induced Apoptosis) mouse, transgene-expressing cells of the myeloid lineage undergo death by apoptosis in the presence of the drug AP20187. Enhanced green fluorescent protein (EGFP) was fused to the suicide gene to allow identification of transgene-expressing cells. Tumor induction was accomplished by subdermal and intravenous injections of B16-F10 melanoma cells. Metastasis in mice with depleted macrophages was compared to metastasis in normal control mice. The lungs and kidneys were examined for metastatic cells. The macrophage-depleted groups showed significantly less metastasis (P>0.001) compared to the control groups. We theorize that macrophages may aid the metastatic process by fusing with melanoma cells. Using appropriate cell markers and fluorescence-activated cell sorting, we were able to detect a small population of double-positive cells. We confirmed cell fusion by microscopic analysis, visualizing the cell's morphology by both immunohistochemistry and immunofluorescence. The presence of double-positive cells suggests macrophage/cancer cell fusion could be a possible mechanism for metastasis.