摘要目的：观察在巨噬细胞（ Mφ）－胶质瘤干细胞（ GSCs ）双色荧光示踪体外共培养模型中，二者间的相互作用，以多指标验证融合细胞的存在及其生物学特性，并分析相关分子机制。方法将红色荧光蛋白基因稳定转染的人GSCs细胞株SU4－RFP，与源于绿色荧光蛋白（ EGFP） Balb／c裸小鼠的Mφ体外共培养，细胞工作站下观察SU4－RFP与Mφ间的包括融合在内的多种相互作用，克隆高增殖力RFP／EGFP双阳性细胞，以免疫印迹、荧光探针原位杂交（ FISH）、免疫细胞化学染色和染色体核型分析多指标鉴定融合细胞，命名为Mφ与GSCs的融合细胞（ F－Mφ），分析其生物学特性及相关分子机制。结果体外共培养发现高增殖力EGFP／RFP双阳性细胞存在，单克隆并连续传代后在转录、翻译及荧光蛋白表达水平均证实RFP和EGFP共表达。融合细胞共表达Mφ标志物CD68和GSCs标志物Nestin，兼具两种细胞特征性染色体，证实其源于SU4－RFP与Mφ的自发融合。融合细胞的增殖速度、侵袭能力均高于SU4－RFP。融合细胞转染miR－146b－5p后，STAT3表达下降，凋亡率上升（18．83％），而致瘤率（4／5）及肿瘤体积（9．7 mm ±1．6 mm）均下降。结论 Mφ可与GSCs自发融合，融合细胞转化后恶性程度更高，与miR－146b－5p下调介导STAT3通路激活相关。

abstractsObjective To observe mutual interactions between macrophages ( Mφ) and glioma stem cells (GSCs)in dual-color tracing model in vitro, to identify the biological characteristics of fusion cells in multiple levels, and to analysis the relevant molecular mechanisms .Methods Red fluorescent protein ( RFP) gene was stably transfected into human GSCs cell line SU 4.Mφcells were obtained from Balb/c nude mice with enhanced green fluorescent protein ( EGFP) expression.Then two cells were co-cultured in dual-color tracing platform.RFP/EGFP double positive cells with high proliferation ability were mono-cloned. The fusion cells were verified by Western blot , fluorescence in situ hybridization , immunocytochemistry and chromosome karyotype analysis .The biological characteristics of fusion cells were further analyzed , together with relevant molecular changes .Results RFP /EGFP double positive cells were obtained through in vitro co-culture.RFP and EGFP coexpression were proved at transcriptional and translational levels in the fusion cells .They also co-expressed GSCs marker Nestin and Mφmarker CD68, and karyotype analysis showed two types of characteristic chromosomes , which confirmed that the fusion cells originated from spontaneous fusion between SU 4-RFP and Mφ.Fusion cell proliferation rate and invasion ability were higher than SU4-RFP, which were relevant with down-regulation of miR-146b-5p and activation of STAT3.Fusion cells transfected with miR-146b-5p showed a higher apoptosis rate (18.83%) and lower tumor formation ( 4/5 ) . Conclusion Mφcould fuse with GSCs spontaneously in local tumor micro-environment.The proliferation and invasion abilities of fusion cells were higher than their parent cells , which were relevant with down-regulation of miR-146b-5p and activation of STAT3. It revealed the possible mechanisms of malignant progression of gliomas .