本文選題：膠質(zhì)瘤干細(xì)胞　切入點(diǎn)：惡性轉(zhuǎn)化　出處：《蘇州大學(xué)》2016年碩士論文

【摘要】：目的觀察膠質(zhì)瘤干細(xì)胞(GSCs)與樹突狀細(xì)胞(DC)體外相互作用,并對膠質(zhì)瘤干細(xì)胞誘導(dǎo)惡性轉(zhuǎn)化的樹突狀細(xì)胞生物學(xué)特征進(jìn)行分析,并比較轉(zhuǎn)化前后的分子差異,為初步探討其分子機(jī)制奠定基礎(chǔ)。方法取表達(dá)綠色熒光蛋白(EGFP)的小鼠骨髓腔沖洗細(xì)胞,從中誘導(dǎo)培養(yǎng)樹突狀細(xì)胞,然后與表達(dá)紅色熒光蛋白(RFP)的人膠質(zhì)瘤干細(xì)胞SU3共培養(yǎng),觀察二者間的相互作用,從中單克隆增殖能力強(qiáng)的共表達(dá)EGFP和RFP雙陽性細(xì)胞,檢測相關(guān)分子標(biāo)志物,并行染色體分析和致瘤試驗(yàn)。在此基礎(chǔ)上,進(jìn)一步分析轉(zhuǎn)化的DC與正常DC基因表達(dá)譜水平的差異。結(jié)果轉(zhuǎn)化的共表達(dá)EGFP和RFP細(xì)胞形態(tài)符合樹突狀細(xì)胞特征,DC標(biāo)志蛋白CD11c、CD80、信號(hào)調(diào)節(jié)蛋白α(SIRP-α)均高表達(dá),此外表達(dá)鼠源性細(xì)胞標(biāo)志物β-肌動(dòng)蛋白;且兼具高增殖、高侵襲力和強(qiáng)致瘤的能力;染色體核型為異倍體,條數(shù)為93±10條。轉(zhuǎn)化后的DC與正常DC相比,全基因組表達(dá)譜數(shù)據(jù)分析顯示,三倍以上差異變化基因3734個(gè),其中上調(diào)2216個(gè),下調(diào)1518個(gè);KEGG通路共涉及72條。其中經(jīng)RT-PCR驗(yàn)證,涉及TAK1、IGF1的AKT-FOXO信號(hào)通路參與到了GSCs誘導(dǎo)的DC惡性轉(zhuǎn)化相關(guān)過程。結(jié)論膠質(zhì)瘤干細(xì)胞SU3-RFP體外可誘導(dǎo)DC發(fā)生惡性轉(zhuǎn)化,惡性轉(zhuǎn)化后的DC保留了樹突狀細(xì)胞的分子標(biāo)記物,比SU3-RFP具有更強(qiáng)的增殖能力和侵襲能力;AKT-FOXO信號(hào)通路與DC的惡性轉(zhuǎn)化相關(guān)。
[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 偽 -SIRP- 偽. 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 鹵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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R739.41

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