摘要
目的研究土木香内酯(ALT)增强胶质瘤干细胞(GSCs)对化疗敏感性的分子机制。方法胶质瘤细胞系U87培养后提取并鉴定GSCs;评价GSCs细胞球细胞的自我更新能力及ALT对其自我更新能力的影响;检测ALT对GSCs中PI3K/Akt信号通路相关蛋白的影响,分析ALT对GSCs干性维持产生的抑制作用与PI3K/Akt信号通路的关系。结果 U87细胞在DMEM培养基中生长方式为单层细胞贴壁生长,24~48 h后干细胞培养液分离培养可见细胞呈球形悬浮状态生长,均可见子细胞球形成;GSCs中干细胞表面标记物CD133^+、Nestin阳性。ALT对GSCs成球能力和表面标志物CD133^+随时间延长下降(P<0.05)。ALT作用24~48 h时,p-PI3K、p-Akt蛋白表达显著下降(P<0.05)。IGF-1组GSCs细胞球个数、p-PI3K、p-Akt、CD133^+与对照组比较差异无统计学意义(P>0.05);ALT组GSCs细胞球个数、p-PI3K、p-Akt、CD133^+均低于对照组(P<0.05);IGF-1组GSCs细胞球个数、p-PI3K、p-Akt、CD133^+均较ALT组显著回升(P<0.05)。结论 ALT对GSCs的干性维持,或与PI3K/Akt信号通路相关,值得临床重视。
Objective To study molecular mechanism of Alantolactone(ALT) in enhancing chemotherapy sensitivity of glioma stem cells(GSCs). Methods GSCs were extracted from glioma cell line U87 after cultivation and identified. Self-renewal ability of GSCs globular cells and effect of ALT on its self-renewal ability were evaluated. Effects of ALT on PI3K/Akt signaling pathway-related proteins in GSCs were detected, and relationship between inhibition effect of ALT on GSCs dry maintenance and PI3K/Akt signaling pathway was analyzed. Results Growth pattern of U87 cells in DMEM culture medium was cell monolayer wall growth. After 24-48 h of isolation and culture, culture medium of stem cells showed spherical suspension growth, and all of them showed formation of daughter-cell ball. Surface markers CD133^+ and Nestin of stem cells in GSCs were positive. Balling ability of ALT to GSCs and surface marker CD133^+ level were significantly decreased with prolonging time(P<0.05). Expressions of p-PI3K and p-Akt proteins were significantly decreased after 24-48 h of ALT treatment(P<0.05). There were no significant differences in number of GSCs globules, p-PI3K, p-Akt and CD133^+ levels between IGF-1 group and control group(P>0.05). Number of GSCs globules, p-PI3K, p-Akt and CD133^+ levels in ALT group were lower than those in control group(P<0.05). Number of GSCs globules, p-PI3K, p-Akt and CD133^+ levels in IGF-1 group were significantly increased compared with those in ALT group(P<0.05). Conclusion GSCs stemness maintenance by ALT may be related to PI3K/Akt signaling pathway, which deserves attention in clinic.
作者
赵勇
朱建国
单显民
王毅东
梁锋
王迅
ZHAO Yong;ZHU Jian-guo;SHAN Xian-min;WANG Yi-dong;LIANG Feng;WANG Xun(Department of Neurosurgery,the Third People's Hospital of Dalian,Dalian,Liaoning 116033,China)
出处
《解放军医药杂志》
CAS
2019年第11期15-19,共5页
Medical & Pharmaceutical Journal of Chinese People’s Liberation Army
基金
2018年辽宁省自然科学基金计划项目(20180827-01)
