紫草素对脑胶质瘤干细胞干性维持的相关研究  

Effects of shikonin on stemness maintance of glioma stem cells

作　　者：刘静[1] 笪祖科[1] 李振[1] 薛一雪[2] 刘丽波[2] 王萍[2] 刘云会[1] 

机构地区：[1]中国医科大学附属盛京医院第一神经外科,辽宁沈阳110004 [2]中国医科大学基础医学院神经生物学教研室,辽宁沈阳110001

出　　处：《中国药理学通报》2016年第1期49-54,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81502179;81372484;81372682;81172197)

摘　　要：目的研究紫草素对脑胶质瘤干细胞(GSCs)干性维持的影响及相关分子机制。方法应用免疫荧光方法对分离提取的细胞球进行干细胞表面标志性分子CD133和nestin的检测;紫草素(2μmol·L-1)处理GSCs 12、24和48 h后,光镜下观察紫草素对GSCs悬浮细胞球形态的影响;采用亚球形成实验评估紫草素对GSCs自我更新能力的影响;应用Western blot方法检测紫草素作用下GSCs干细胞标志分子CD133的表达变化,同时检测紫草素作用下,GSCs中PI3K、p-PI3K、Akt和p-Akt的表达变化;联合应用胰岛素样生长因子-1(IGF-1)后检测紫草素作用下,GSCs干性维持的改变。结果分离提取的细胞球表面CD133和nestin呈阳性表达;紫草素能够明显抑制GSCs悬浮细胞球的形态和二代细胞球的形成,同时能够降低CD133的表达;与对照组相比,紫草素作用下,GSCs中PI3K和Akt的表达无变化,pPI3K和p-Akt的表达呈现药物时间依赖性明显下降;此外,IGF-1能够明显改善紫草素对GSCs干性维持的抑制作用。结论紫草素能够明显抑制GSCs的干性维持,其机制可能与PI3K/Akt信号通路有关。Aim To explore the effect of shikonin on stemness maintance of glioma stem cells( GSCs).Methods After the U87-MG cells were cultured and isolated,the sphere cells were identified by immunofluorescent staining. The alteration of stemness of GSCs by shikonin treatment( 2 μmol · L- 1) for 12 h,24 h and 48 h was valued by morphological detection using optical microscope and sub-sphere forming assay. Moreover,the related markers of stem cells, such as CD133,were detected in shikonin treated GSCs by western blot assay. Protein expression of PI3 K, pPI3 K,Akt and p-Akt was detected by western blot after shikonin treatment alone. Furthermore,by combination with insulin-like growth factor-1( IGF-1),we observed the alteration of stemness maintance of shikonin-treated GSCs. Results The presence of neural stem cell related markers CD133 and nestin proved the characteristics of GSCs. Shikonin treatment significantly inhibited the morphology of GSCs and the sub-sphere forming. Besides,the reduced expression of CD133 was detected in shikonin treated GSCs. Though,the expression of PI3 K and Akt did not change compared with the control group,the expression of p-PI3 K and p-Akt was reduced. Furthermore,the combination of IGF-1 markedly attenuated the inhibitory effect of shikonin on stemness maintance of GSCs. Conclusion The stemness maintance of GSCs can be significantly inhibited by shikonin treatment,in which PI3 K / Akt pathway is involved.

关 键 词：紫草素 胶质瘤干细胞 干性维持 PI3K/AKT信号通路 胰岛素样生长因子-1 自我更新能力 

分 类 号：R284.4[医药卫生—中药学] R329.24[医药卫生—中医学]