对顺铂耐药的骨肉瘤细胞干细胞特性变化及其与Notch信号通路的关系  

Changes of stem cell properties in cisplatin-resistant osteosarcoma cells and its relationship with Notch signaling pathway

作　　者：李提 方硕[1] 余铃[1] 杨俭[1] 郭卫春[1] 

机构地区：[1]武汉大学人民医院,武汉430060

出　　处：《山东医药》2016年第48期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81341078)

摘　　要：目的探讨对顺铂耐药的骨肉瘤细胞干细胞特性变化及其与Notch信号通路的关系。方法取对数生长期的对顺铂耐药的人骨肉瘤143B-TK细胞作为观察组,以未经顺铂作用的人骨肉瘤143B-TK细胞作为对照组。采用实时荧光定量PCR法检测两组干细胞相关基因(TERT、Sox2、Oct4、NANOG)、Notch信号通路相关基因(Hey L、Hey2、Hey1、Hes5、Hes1)mRNA相对表达量;两组加入或不加入Notch信号通路特异性抑制剂DAPT,采用悬浮克隆球试验检测成球能力(以悬浮克隆球数量表示)。结果两组NANOG、Hey2 mRNA相对表达量比较差异均无统计学意义(P均>0.05)。观察组TERT、Sox2、Oct4、Hey L、Hey1、Hes5、Hes1 mRNA相对表达量均高于对照组(P<0.05或<0.01)。不加入DAPT的观察组悬浮克隆球数量明显多于加入DAPT的观察组和不加入DAPT的对照组(P均<0.01)。结论对顺铂耐药的骨肉瘤细胞干细胞特性增强;Notch信号通路激活可能是导致其干细胞特性增强和细胞耐药的主要原因。Objective To investigate the characteristics of cisplatin-resistant osteosarcoma stem cells and its relation- ship with Notch signaling pathway. Methods The cisplatin-resistant human osteosarcoma 143B-TK ceils in the logarithmic growth phase （observation group） were selected, and 143B-TK ceils without the treatment of eisplatin were taken as the control group. Real-time quantitative PCR was used to detect the expression of the stem cells-related genes （TERT, Sox2, Oct4 and NANOG）, and the Notch signaling pathway-related genes （HeyL, Hey2, Heyl, HesS, and Hesl ） in the two groups. Cell forming ability was detected by the formation of sphere number in the two groups with or without adding DAPT. Results No significant difference was found in the relative expression of NANOG and Hey2 between the two groups （ all P 〉 0.05）. The relative expression of TERT, Sox2, Oct4, HeyL, Heyl, Hes5, and Hesl in the observation group was higher than that in the control group （P 〈0. 05 or P 〈0.01 ）. The sphere formation number in the observation group without adding DAPT was more than that in the observation group with adding DAFF and that in the control group without adding D APT （ all P 〈0.01 ）. Conclusion The stem cell properties of cisplatin-resistant osteosarcoma cells enhance, and the acti- vation of Notch signaling pathway may be main reason for the enhancement of stem cell properties and drug resistance.

关 键 词：骨肉瘤 顺铂耐药 肿瘤干细胞 NOTCH信号通路 

分 类 号：R457.7[医药卫生—治疗学]