胶质瘤干细胞诱导树突状细胞恶性转化及相关分子机制的初步研究

发布时间：2018-03-28 22:28

本文选题：胶质瘤干细胞　切入点：恶性转化　出处：《苏州大学》2016年硕士论文

【摘要】：目的观察胶质瘤干细胞(GSCs)与树突状细胞(DC)体外相互作用,并对胶质瘤干细胞诱导恶性转化的树突状细胞生物学特征进行分析,并比较转化前后的分子差异,为初步探讨其分子机制奠定基础。方法取表达绿色荧光蛋白(EGFP)的小鼠骨髓腔冲洗细胞,从中诱导培养树突状细胞,然后与表达红色荧光蛋白(RFP)的人胶质瘤干细胞SU3共培养,观察二者间的相互作用,从中单克隆增殖能力强的共表达EGFP和RFP双阳性细胞,检测相关分子标志物,并行染色体分析和致瘤试验。在此基础上,进一步分析转化的DC与正常DC基因表达谱水平的差异。结果转化的共表达EGFP和RFP细胞形态符合树突状细胞特征,DC标志蛋白CD11c、CD80、信号调节蛋白α(SIRP-α)均高表达,此外表达鼠源性细胞标志物β-肌动蛋白;且兼具高增殖、高侵袭力和强致瘤的能力;染色体核型为异倍体,条数为93±10条。转化后的DC与正常DC相比,全基因组表达谱数据分析显示,三倍以上差异变化基因3734个,其中上调2216个,下调1518个;KEGG通路共涉及72条。其中经RT-PCR验证,涉及TAK1、IGF1的AKT-FOXO信号通路参与到了GSCs诱导的DC恶性转化相关过程。结论胶质瘤干细胞SU3-RFP体外可诱导DC发生恶性转化,恶性转化后的DC保留了树突状细胞的分子标记物,比SU3-RFP具有更强的增殖能力和侵袭能力;AKT-FOXO信号通路与DC的恶性转化相关。
[Abstract]:Objective to observe the interaction between glioma stem cells (GSCs) and dendritic cells (DC) in vitro, and to analyze the biological characteristics of malignant transformation induced by glioma stem cells, and to compare the molecular differences before and after transformation. Methods Murine myelin washing cells expressing green fluorescent protein (EGFP) were used to induce the culture of dendritic cells, and then co-cultured with human glioma stem cells (SU3) expressing red fluorescent protein. We observed the interaction between the two cells and coexpressed EGFP and RFP double positive cells with strong proliferation ability, detected the related molecular markers, and analyzed the chromosomes and tumorigenic test. The difference of gene expression profile between transformed DC and normal DC was further analyzed. Results the morphology of transformed EGFP and RFP cells was in line with dendritic cell characteristic DC marker CD11cnc-CD80 and signal regulated protein 伪 -SIRP- 伪. In addition, 尾 -actin was expressed as a cell marker of murine origin, and had the ability of high proliferation, high invasiveness and strong tumorigenesis. The chromosome karyotype was aneuploidy and the number of strips was 93 卤10. The transformed DC was compared with normal DC. The whole genome expression profile data showed that 3734 genes were more than three times different, including 2216 up-regulated genes and 72 down-regulated 1518 KEGG transduction pathways, which were verified by RT-PCR. The AKT-FOXO signaling pathway involved in TAK1-IGF1 is involved in the process of DC malignant transformation induced by GSCs. Conclusion Glioma stem cell SU3-RFP can induce DC malignant transformation in vitro, and the DC after malignant transformation retains the molecular marker of dendritic cells. Compared with SU3-RFP, AKT-FOXO signaling pathway has stronger proliferative and invasive ability and is associated with malignant transformation of DC.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R739.41

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