DAPT和PI-103对人胶质瘤干细胞侵袭、迁移能力的影响  

Effects of DAPT combined with PI-103 on invasion and migration of human glioma stem cells

作　　者：周星辰 闵敬亮 束汉生[1] 王大巍 程哲 张辉[1] 王昊[1] 杨光 ZHOU Xingchen;MIN Jingliang;SHU Hansheng;WANG Dawei;CHENG Zhe;ZHANG Hui;WANG Hao;YANG Guang(The Second Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第二附属医院,安徽蚌埠233000

出　　处：《山东医药》2020年第24期9-11,共3页Shandong Medical Journal

基　　金：蚌埠医学院自然科学基金面上项目(BYKY18143);安徽省青年自然科学基金(1908085QH334)。

摘　　要：目的观察联合应用Notch1信号通路抑制剂(DAPT)和PI3K/Akt/mTOR信号通路抑制剂(PI-103)对人胶质瘤干细胞(GSCs)侵袭、迁移能力的影响。方法采用免疫磁珠法从U87细胞中分选获得GSCs;取GSCs分为DAPT处理组(A组)、PI-103处理组(B组)、DAPT和PI-103共同处理组(C组)、药物溶媒对照组(D组),A组加入20μmol/L的DAPT培养,B组加入4μmol/L的PI-103培养,C组加入20μmol/L的DAPT+4μmol/L的PI-103共培养,D组不作任何处理。Western blotting法检测各组Notch1活化标志物(Hes1、pmTOR蛋白),Transwell侵袭和迁移实验观察各组细胞侵袭、迁移能力(穿膜细胞数)。结果与D组和A组相比,B、C组Hes1蛋白相对表达量降低(P均<0.01);与D组相比,A、B、C组pmTOR蛋白相对表达量降低(P均<0.05);与A、B组比较,C组pmTOR蛋白相对表达量降低(P均<0.05)。与D组相比,A、B、C组侵袭、迁移穿膜细胞数降低(P均<0.05);与A、B组比较,C组侵袭、迁移穿膜细胞数降低(P均<0.05)。结论DAPT和PI-103联合使用可显著降低GSCs的侵袭、迁移能力。Objective To observe the effects of combined application of Notch1 signaling pathway inhibitor(DAPT)and PI3 K/Akt/mTOR signaling pathway inhibitor(PI-103)on the invasion and migration of human glioma stem cells(GSCs).Methods Immunomagnetic bead method was used to obtain GSCs from U87 cells;GSCs were divided into the drug vehicle control group(group D),DAPT treatment group(group A),and PI-103 treatment group(group B),and DAPT and PI-103 co-treatment group(group C).GSCs in the group A were treated with 20μmol/L DAPT,group B with 4μmol/L PI-103,and group C with 20μmol/L DAPT+4μmol/L PI-103,but the group A was not treated.Western blotting was used to detect the invasion and migration of Notch1 activation markers(Hes1 and pmTOR protein),and Transwell invasion and migration assays were used to observe the cell invasion and migration abilities(the number of transmembrane cells)in each group.Results Compared with the control group and group A,the relative expression of Hes1 protein in the groups B and C decreased(F=66.418,P<0.01);compared with the control group,the relative expression of pmTOR protein in the groups A,B,and C decreased(all P<0.05);compared with groups A and B,the relative expression of pmTOR protein in the group C decreased(P<0.05).Compared with the group D,the number of invasive and migrating transmembrane cells in the groups A,B,and C decreased(all P<0.05);compared with groups A and B,the number of invasive and migrating transmembrane cells in the group C decreased(P<0.05).Conclusion The combined use of DAPT and PI-103 can significantly reduce the invasion and migration abilities of GSCs.

关 键 词：胶质瘤 人胶质瘤干细胞 Notch1信号通路 PI3K/Akt/mTOR信号通路 细胞侵袭能力 细胞迁移能力 

分 类 号：R739.41[医药卫生—肿瘤]