摘要
目的探讨双氢青蒿素(dihydroartemisinine,DHA)对人神经胶质瘤生长的影响及机制。方法人胶质瘤U251细胞株传代培养,设置空白对照组、二甲基亚砜(DMSO)组、DAPT组和DHA(10、20、40、80μmol/L)实验组,应用CCK-8和FCM检测细胞增殖、周期及凋亡,Western blot检测Notch1的胞内区域(NICD-1)及其下游靶蛋白Hes1的表达水平。建立裸鼠皮下胶质瘤模型(n=20),按随机数字表法分为DMSO组、DAPT组、DHA(50、100 mg/kg)治疗组,每组5只,根据时间体积曲线计算各组抑瘤率,免疫组化法检测裸鼠皮下移植瘤组织中NICD-1表达水平变化。结果与空白对照组和DMSO组相比,实验组U251细胞增殖率显著下降,G1期细胞所占百分比显著上升,S期细胞所占百分比显著下降,早期凋亡率显著上升(P<0.05),NICD-1及Hes1蛋白表达水平显著低于空白对照组(P<0.05),均呈浓度依赖关系;DHA各治疗组的裸鼠皮下移植瘤生长速率及移植瘤组织内NICD-1表达显著低于DMSO对照组(P<0.05)。结论 DHA有效抑制人神经胶质瘤的生长,其作用机制可能与通过下调NICD-1表达抑制Notch信号通路有关。
Objective To investigate the effects of dihydroartemisinine（ DHA） on the growth of human glioma xenografts in nude mice and explore the underlying mechanism. Methods Human glioma U251 cells were divided into blank control group,DMSO group,DAPT group,and DHA（ 10,20,40,and80 μmol/L） treatment groups. The viability,cell cycle and apoptosis of U251 cells in different groups were analyzed using CCK-8 assay and flow cytometry,and the expression levels of Notch intracellular domain（ NICD-1） and hairy enhance of split 1（ Hes1） in the cells were detected with Western blotting. Twenty nude mice bearing subcutaneous U251 cell xenograft were randomized into DMSO group,DAPT group and DHA（ 50 and 100 mg/kg） treatment groups（ n = 5） for corresponding treatments,and the time-volume curve of tumor growth and the tumor inhibitory rates were calculated; NICD-1 expression in the xenografted tumor tissues were detected with immunohistochemistry. Results Compared with the blank control group and DMSO-treated cells,U251 cells with DHA treatment showed significantly suppressed proliferation,increased percentage of cells in G1 phase,decreased percentage of cells in S phase,and significantly increased early apoptotic rate（ P〈0. 05）. The expression levels of protein NICD-1 and Hes1 were significantly lower in DHA-treated cells than in the blank control cells（ P〈0. 05）; all these effects of DHA exhibited a dose dependence. Compared with DMSO treatment,DHA treatment of the tumor-bearing mice resulted in a significant growth inhibition of the xenografted tumors and lowered NICD-1 expression in the tumor tissues（ P〈0. 05）. Conclusion DHA can effectively inhibit the growth of human glioma xenografts in nude mice possibly by down-regulating NICD-1 expression to inhibit Notch signaling pathway.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2017年第14期1445-1451,共7页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(81371674
81471676)~~
