【摘要】：目的:骨肉瘤干細(xì)胞具有化療耐藥性。本文擬探討耐阿霉素細(xì)胞干細(xì)胞樣特性的改變,以及Notch通路在其中的調(diào)控作用。方法:采用2μM的阿霉素處理骨肉瘤細(xì)胞143B 24 h,去藥繼續(xù)培養(yǎng)5 d,檢測(cè)干細(xì)胞樣特性的改變,包括形態(tài)學(xué)的改變、Stro-1~+/CD117~+雙陽性細(xì)胞比例、干細(xì)胞相關(guān)基因表達(dá)、懸浮成球的能力、EMT特性。qPCR及Western blot檢測(cè)Notch通路受體及靶基因表達(dá)情況。利用Notch抑制劑DAPT預(yù)處理,檢測(cè)其對(duì)耐阿霉素骨肉瘤細(xì)胞的干細(xì)胞樣特性的影響。構(gòu)建裸鼠移植瘤模型,檢測(cè)Notch抑制劑對(duì)體內(nèi)成瘤的影響。結(jié)果:耐阿霉素骨肉瘤細(xì)胞中Stro-1~+/CD117~+比例增高,干細(xì)胞相關(guān)基因Oct4、Sox2表達(dá)量增加,懸浮成球能力增強(qiáng),EMT特性上調(diào)。q PCR及Western blot結(jié)果顯示阿霉素耐藥的骨肉瘤細(xì)胞中Notch受體胞內(nèi)段NICD1及靶基因Hes1、Hey1等表達(dá)量上調(diào)。Notch信號(hào)抑制劑能夠增強(qiáng)骨肉瘤對(duì)阿霉素的化療敏感性,抑制體外阿霉素對(duì)骨肉瘤干細(xì)胞的富集作用。動(dòng)物實(shí)驗(yàn)表明,Notch抑制劑DAPT能夠抑制體內(nèi)成瘤。結(jié)論:阿霉素能夠富集骨肉瘤干細(xì)胞,Notch信號(hào)通路參與其中調(diào)控機(jī)制,抑制Notch通路能夠靶向殺傷骨肉瘤細(xì)胞,增加化療藥物敏感性。
[Abstract]:Objective: osteosarcoma stem cells have chemotherapeutic resistance. The purpose of this study was to investigate the change of stem cell-like characteristics of adriamycin-resistant cells and the regulation of Notch pathway. Methods: osteosarcoma cell line 143B was treated with 2 渭 M adriamycin for 24 h, then cultured for 5 days. The changes of stem cell like characteristics, including morphological changes, ratio of Stro-1 ~ / CD117~ double positive cells and expression of stem cell related genes, were detected. The ability of suspending pelletizing. QPCR and Western blot were used to detect the expression of Notch pathway receptor and target gene. The effects of Notch inhibitor DAPT pretreatment on stem cell-like characteristics of adriamycin-resistant osteosarcoma cells were detected. To study the effect of Notch inhibitor on tumor formation in nude mice. Results: the ratio of Stro-1~ / CD 117 ~ in adriamycin resistant osteosarcoma cells was increased, and the Oct4,Sox2 expression of stem cell related gene was increased. The expression of Notch receptor NICD1 and target gene Hes1,Hey1 were up-regulated in osteosarcoma cells with adriamycin resistance. The results showed that Notch signaling inhibitor could enhance the chemosensitivity of osteosarcoma to doxorubicin, and enhance the chemosensitivity of osteosarcoma to doxorubicin. To inhibit the enrichment of osteosarcoma stem cells by adriamycin in vitro. Animal experiments have shown that DAPT, an inhibitor of Notch, can inhibit tumorigenesis in vivo. Conclusion: doxorubicin can enrich the Notch signaling pathway of osteosarcoma stem cells, and inhibit Notch pathway can target osteosarcoma cells and increase chemosensitivity.
【作者單位】： 武漢大學(xué)人民醫(yī)院骨1科;北京大學(xué)腫瘤醫(yī)院暨北京市腫瘤防治研究所骨與軟組織腫瘤科惡性腫瘤發(fā)病機(jī)制及轉(zhuǎn)化研究教育部重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金(編號(hào):81502575) 中央高校基本科研業(yè)務(wù)費(fèi)專項(xiàng)資金(編號(hào):2042015kf0069)資助~~
【分類號(hào)】：R738

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