胆碱激酶α通过外泌体介导胶质母细胞瘤U87MG细胞的恶性表型  

Choline kinase α mediates the malignant phenotype of glioblastoma U87MG cells through exosomes

作　　者：李嘉麟 邹有瑞[1] 刘阳[1] 赵阳 马辉[1] LI Jialin;ZOU Yourui;LIU Yang;ZHAO Yang;MA Hui(General Hospital of Ningxia Medical University,Ningxia Yinchuan 750004,China.)

机构地区：[1]宁夏医科大学总医院,宁夏银川750004

出　　处：《现代肿瘤医学》2024年第15期2738-2745,共8页Journal of Modern Oncology

基　　金：宁夏自然科学基金项目(编号:2022AAC03559)。

摘　　要：目的:探究胆碱激酶α(Choline kinase alpha,CHKA)通过外泌体影响胶质母细胞瘤U87MG细胞增殖、迁移及侵袭能力的机制。方法:在胶质瘤U87MG细胞中转染shCHKA及其空载对照;采用外泌体分泌抑制剂GW4869抑制外泌体分泌;利用透射电镜及纳米粒径示踪分析鉴定外泌体形态和大小,BCA法鉴定外泌体浓度,Western Blot检测外泌体标志性蛋白表达水平;提取CHKA不同表达组细胞培养上清中的外泌体与U87MG共孵育,CCK-8检测细胞增殖能力,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力。最后,用U87MG细胞构建裸鼠体内模型,观察不同CHKA表达U87MG细胞来源的外泌体对裸鼠成瘤能力的影响。结果:与空载对照细胞相比,shCHKA组细胞培养上清中的外泌体浓度降低(P<0.001),外泌体特异性蛋白的表达水平降低(P<0.001)。加入GW4869后,胶质瘤U87MG细胞的增殖、迁移及侵袭能力均降低(P<0.05,P<0.01,P<0.01)。GW4869可以明显抑制空载对照细胞的增殖、迁移及侵袭能力(P<0.05,P<0.001,P<0.001),但对shCHKA细胞的抑制作用不明显。与加入shNC组外泌体的U87MG细胞相比,加入shCHKA组外泌体的U87MG细胞的增殖、迁移、侵袭能力明显减弱(P<0.01,P<0.05,P<0.001)。裸鼠体内成瘤实验表明,与注射shNC组外泌体相比,注射shCHKA组外泌的裸鼠移植瘤生长能力明显减弱(P<0.01)。结论:CHKA的表达可能通过外泌体介导胶质母细胞瘤U87MG细胞的恶性表型。Objective:To investigate the effect of Choline kinase alpha(CHKA)on the proliferation,migration and invasion of glioblastoma U87MG cells through exosomes.Methods:U87MG cells were transfected with shCHKA and its empty vector control.Exosome secretion was inhibited by exosome secretion inhibitor GW4869.Transmission electron microscopy and nanoparticle size tracing analysis were used to identify the morphology and size of exosomes.BCA method was used to identify the concentration of exosomes,and Western Blot was used to detect the expression levels of exosome marker proteins.Exosomes extracted from the cell culture supernatant of different CHKA expression groups were co-incubated with U87MG.CCK-8 assay was used to detect cell proliferation,scratch test was used to detect cell migration,and Transwell test was used to detect cell invasion.Finally,U87MG cells were used to construct a nude mouse model to observe the effect of exosomes derived from U87MG cells with different CHKA expressions on the tumorigenic ability of nude mice.Results:Compared with the empty vector control cells,the concentration of exosomes in the culture supernatant of shCHKA cells was decreased(P<0.001),and the expression level of exosome-specific proteins was decreased ( P <0.001).After adding GW4869,the proliferation,migration and invasion ability of glioma U87MG cells were decreased ( P <0.05, P <0.01, P <0.01).GW4869 significantly inhibited the proliferation,migration and invasion of the empty vector control cells ( P <0.05, P <0.001, P <0.001),but the inhibitory effect on shCHKA cells was not obvious.Compared with U87MG cells added with exosomes from shNC group,the proliferation,migration,and invasion abilities of U87MG cells added with exosomes from shCHKA group were significantly weakened ( P <0.01, P <0.05, P <0.001).The tumor formation experiment in nude mice showed that the tumor growth ability of nude mice injected with exosomes from the shCHKA group was significantly reduced compared with that of the shNC group ( P <0.01). Conclusion:

关 键 词：胆碱激酶α 外泌体 胶质母细胞瘤 增殖 迁移 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]