阿霉素通过激活Notch信号通路促进骨肉瘤细胞干性特性
发布时间：2018-08-24 07:54
【摘要】：目的:骨肉瘤干细胞具有化疗耐药性。本文拟探讨耐阿霉素细胞干细胞样特性的改变,以及Notch通路在其中的调控作用。方法:采用2μM的阿霉素处理骨肉瘤细胞143B 24 h,去药继续培养5 d,检测干细胞样特性的改变,包括形态学的改变、Stro-1~+/CD117~+双阳性细胞比例、干细胞相关基因表达、悬浮成球的能力、EMT特性。qPCR及Western blot检测Notch通路受体及靶基因表达情况。利用Notch抑制剂DAPT预处理,检测其对耐阿霉素骨肉瘤细胞的干细胞样特性的影响。构建裸鼠移植瘤模型,检测Notch抑制剂对体内成瘤的影响。结果:耐阿霉素骨肉瘤细胞中Stro-1~+/CD117~+比例增高,干细胞相关基因Oct4、Sox2表达量增加,悬浮成球能力增强,EMT特性上调。q PCR及Western blot结果显示阿霉素耐药的骨肉瘤细胞中Notch受体胞内段NICD1及靶基因Hes1、Hey1等表达量上调。Notch信号抑制剂能够增强骨肉瘤对阿霉素的化疗敏感性,抑制体外阿霉素对骨肉瘤干细胞的富集作用。动物实验表明,Notch抑制剂DAPT能够抑制体内成瘤。结论:阿霉素能够富集骨肉瘤干细胞,Notch信号通路参与其中调控机制,抑制Notch通路能够靶向杀伤骨肉瘤细胞,增加化疗药物敏感性。
[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.
【作者单位】： 武汉大学人民医院骨1科;北京大学肿瘤医院暨北京市肿瘤防治研究所骨与软组织肿瘤科恶性肿瘤发病机制及转化研究教育部重点实验室;
【基金】：国家自然科学基金(编号:81502575) 中央高校基本科研业务费专项资金(编号:2042015kf0069)资助~~
【分类号】：R738
[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.
【作者单位】： 武汉大学人民医院骨1科;北京大学肿瘤医院暨北京市肿瘤防治研究所骨与软组织肿瘤科恶性肿瘤发病机制及转化研究教育部重点实验室;
【基金】：国家自然科学基金(编号:81502575) 中央高校基本科研业务费专项资金(编号:2042015kf0069)资助~~
【分类号】：R738
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