背景:异种移植瘤中宿主来源间质细胞恶性转化已有报道,但是对恶变的机制知之甚少。目的:旨在使用双色荧光示踪的体内模型研究宿主来源肿瘤间质细胞的恶性转化,并探讨可能的机制。方法:将稳定转染红色荧光蛋白基因的人脑胶质瘤起始细胞(g...背景:异种移植瘤中宿主来源间质细胞恶性转化已有报道,但是对恶变的机制知之甚少。目的:旨在使用双色荧光示踪的体内模型研究宿主来源肿瘤间质细胞的恶性转化,并探讨可能的机制。方法:将稳定转染红色荧光蛋白基因的人脑胶质瘤起始细胞(glioma-initiatingcellstransfectedwithred fluorescent protein gene,GICs-RFP)接种到表达绿光荧光蛋白的裸鼠体内,建立具有2种荧光的动物模型;通过激光共聚焦观察移植瘤的荧光表达;流式细胞仪检测和分选各种荧光细胞,包括绿色荧光蛋白细胞、红色荧光蛋白细胞和表达这2种荧光的融合细胞;体外亚克隆得到具有恶性肿瘤细胞特征的同时表达绿色荧光蛋白/红色荧光蛋白两种荧光的融合细胞;体外鉴定细胞来源、表面标记物和恶性特征,裸鼠皮下移植验证融合细胞致瘤特性。实验经苏州大学动物实验伦理委员会批准,批准号为ECSU-201800090。结果与结论:GICs-RFP在绿色荧光蛋白裸小鼠的体内致瘤率均为100%(14/14);动物移植瘤模型中均可观察到融合细胞,分选、克隆到的融合细胞不仅共表达绿色荧光蛋白和红色荧光蛋白,而且共表达GICs-RFP标记物Nestin和骨髓间充质干细胞标记物CD44、CD105和CD29;融合细胞在体外表现出高度增殖活性,较GICs-RFP更具侵袭和迁移特征;融合细胞(1×105个/只)在无胸腺裸小鼠的致瘤率为100%(4/4)。提示GICs-RFP自发融合宿主骨髓间充质干细胞并诱导恶性转化,为肿瘤异质性的细胞来源提供了新的双色荧光示踪的证据。展开更多
Objective: Real-time monitoring of cytokine secretion at the single immunocyte level,based on the concept of immune cells, sociology has been recently reported. However,the relationships between glioma-initiating cell...Objective: Real-time monitoring of cytokine secretion at the single immunocyte level,based on the concept of immune cells, sociology has been recently reported. However,the relationships between glioma-initiating cells(GICs) and host immune cells and their mutual interactions in the tumor microenvironment have not been directly observed and remain unclear. Methods: The dual fluorescence tracing technique was applied to label the co-cultured GICs and host macrophages(M?), and the interactions between the two types of cells were observed using a live cell imaging system. Fusion cells in the co-culture system were monocloned and proliferated in vitro and their social interactions were observed and recorded. Results: Using real-time dynamic observation of target cells, 6 types of intercellular conjunction microtubes were found to function in the transfer of intercellular information between GICs and M?; GICs and host M? can fuse into hybrid cells after several rounds of mutual interactions, and then these fusion cells fused with each other; Fusion cells generated offspring cells through symmetrical and asymmetrical division or underwent apoptosis. A "cell in cell" phenomenon was observed in the fusion cells, which was often followed by cell release, namely entosis. Conclusions: Preliminary studies revealed the patterns of cell conjunction via microtubes between GICs and host M? and the processes of cell fusion, division, and entosis. The results revealed malignant transformation of host M?, induced by GICs, suggesting complex social relationships among tumor-immune cells in gliomas.展开更多
文摘背景:异种移植瘤中宿主来源间质细胞恶性转化已有报道,但是对恶变的机制知之甚少。目的:旨在使用双色荧光示踪的体内模型研究宿主来源肿瘤间质细胞的恶性转化,并探讨可能的机制。方法:将稳定转染红色荧光蛋白基因的人脑胶质瘤起始细胞(glioma-initiatingcellstransfectedwithred fluorescent protein gene,GICs-RFP)接种到表达绿光荧光蛋白的裸鼠体内,建立具有2种荧光的动物模型;通过激光共聚焦观察移植瘤的荧光表达;流式细胞仪检测和分选各种荧光细胞,包括绿色荧光蛋白细胞、红色荧光蛋白细胞和表达这2种荧光的融合细胞;体外亚克隆得到具有恶性肿瘤细胞特征的同时表达绿色荧光蛋白/红色荧光蛋白两种荧光的融合细胞;体外鉴定细胞来源、表面标记物和恶性特征,裸鼠皮下移植验证融合细胞致瘤特性。实验经苏州大学动物实验伦理委员会批准,批准号为ECSU-201800090。结果与结论:GICs-RFP在绿色荧光蛋白裸小鼠的体内致瘤率均为100%(14/14);动物移植瘤模型中均可观察到融合细胞,分选、克隆到的融合细胞不仅共表达绿色荧光蛋白和红色荧光蛋白,而且共表达GICs-RFP标记物Nestin和骨髓间充质干细胞标记物CD44、CD105和CD29;融合细胞在体外表现出高度增殖活性,较GICs-RFP更具侵袭和迁移特征;融合细胞(1×105个/只)在无胸腺裸小鼠的致瘤率为100%(4/4)。提示GICs-RFP自发融合宿主骨髓间充质干细胞并诱导恶性转化,为肿瘤异质性的细胞来源提供了新的双色荧光示踪的证据。
基金Supported by the National Natural Science Foundation of China(Grant No.81472739)the Natural Science Foundation of Jiangsu Province(Grant No.BK20151214)
文摘Objective: Real-time monitoring of cytokine secretion at the single immunocyte level,based on the concept of immune cells, sociology has been recently reported. However,the relationships between glioma-initiating cells(GICs) and host immune cells and their mutual interactions in the tumor microenvironment have not been directly observed and remain unclear. Methods: The dual fluorescence tracing technique was applied to label the co-cultured GICs and host macrophages(M?), and the interactions between the two types of cells were observed using a live cell imaging system. Fusion cells in the co-culture system were monocloned and proliferated in vitro and their social interactions were observed and recorded. Results: Using real-time dynamic observation of target cells, 6 types of intercellular conjunction microtubes were found to function in the transfer of intercellular information between GICs and M?; GICs and host M? can fuse into hybrid cells after several rounds of mutual interactions, and then these fusion cells fused with each other; Fusion cells generated offspring cells through symmetrical and asymmetrical division or underwent apoptosis. A "cell in cell" phenomenon was observed in the fusion cells, which was often followed by cell release, namely entosis. Conclusions: Preliminary studies revealed the patterns of cell conjunction via microtubes between GICs and host M? and the processes of cell fusion, division, and entosis. The results revealed malignant transformation of host M?, induced by GICs, suggesting complex social relationships among tumor-immune cells in gliomas.
