转移相关蛋白3通过活性氧参与胶质瘤耐药的分子机制研究  

Molecular mechanism of metastasis-associated protein 3 involvement in glioma drug resistance through reactive oxygen

作　　者：张应杰 吴再辉 李晓龙 李林 ZHANG Yingjie;WU Zaihui;LI Xiaolong;LI Lin(Neurosurgery Department,Tangshan Hongci Hospital,Tangshan,Hebei 063000,P.R.China;Neurosurgery Department,Shijiazhuang Great Wall Integrated Traditional Chinese and Western Medicine Hospital,Shijiazhuang,Hebei 050000,P.R.China;Neurosurgery Department,North China University of Science and Technology Affiliated Hospital,Tangshan,Hebei 063000,P.R.China)

机构地区：[1]唐山弘慈医院神经外科,河北唐山063000 [2]石家庄长城中西医结合医院神经外科,石家庄050000 [3]华北理工大学附属医院神经外科,河北唐山063000

出　　处：《华西医学》2024年第5期732-738,共7页West China Medical Journal

基　　金：河北省卫生健康委员会科研项目(20210338)。

摘　　要：目的 探讨转移相关蛋白3(metastasis-associated protein 3, MTA3)通过活性氧参与胶质瘤耐药的分子机制。方法 采用蛋白印迹检测胶质瘤干细胞(glioma stem cells, GSC)、非GSC的表达。其中,GSC包括U87和SHG44细胞,非GSC包括U87s和SU-2细胞。过表达MTA3后,将U87和SHG44细胞分为Lv-scr组和Lv-MTA3组,通过神经球形成试验测试胶质瘤细胞的自我更新能力,在常氧条件或缺氧条件下暴露于0、2、4、6、8、10 Gy X线照射后检查细胞存活分数。采用流式细胞术分析检测细胞凋亡和检测活性氧表达,对干细胞标志物CD133和巢蛋白以及分化标志物胶质纤维酸性蛋白(星形胶质细胞)和神经元类型Ⅲβ-微管蛋白克隆进行免疫荧光染色。结果 在GSC中,U87s和SU-2组MTA3表达较低;过表达MTA3后,Lv-MTA3在U87s和SU-2中表达较Lv-scr组升高。在常氧或缺氧条件下,U87和SU-2分别比胶质瘤细胞系U87和SHG44更具放射抗性。与非GSC比较,GSC基底活性氧形成减少,非GSC活性氧生成增加。在常氧或缺氧条件下暴露于不同剂量的X线后,具有过表达MTA3的GSC比具有稳定整合的GSC更具有放射敏感性。此外,MTA3过表达导致GSC中的OER略高。MTA3过表达降低了2种干细胞系的CD133和巢蛋白免疫抑素,并增加了神经胶质纤维酸性蛋白和神经元类型Ⅲβ-微管蛋白克隆免疫染色,差异有统计学意义(P<0.05)。结论 MTA3在GSC中下调,MTA3过表达降低了GSC在体外和体内的耐辐射性和干性。MTA3在通过活性氧调节GSC的放射敏感性和干性方面起重要作用。Objective To investigate the molecular mechanism by which metastasis-associated protein 3(MTA3)participates in glioma resistance through reactive oxygen species.Methods Protein expression in glioma stem cells(GSCs)and non-GSCs was detected using Western blotting.GSCs included U87 and SHG44 cells,while non-GSCs included U87s and SU-2 cells.After overexpressing MTA3,U87 and SHG44 cells were divided into Lv-scr and Lv-MTA3 groups.The self-renewal capacity of glioma cells was assessed through a neurosphere formation assay.Cell survival fractions were examined following exposure to 0,2,4,6,8,and 10 Gy X-ray irradiation under normoxic or hypoxic conditions.Apoptosis and reactive oxygen species expression were analyzed using flow cytometry.Immunofluorescence staining was performed to detect the stem cell markers CD133 and nestin,as well as the differentiation markers glial fibrillary acidic protein(GFAP,for astrocytes)and neuronal classⅢβ-tubulin.Results In GSCs,MTA3 expression was lower in the U87s and SU-2 groups. After MTA3 overexpression, Lv-MTA3 expression was higher in U87s and SU-2compared to the Lv-scr group. Under normoxic or hypoxic conditions, U87 and SU-2 showed greater radioresistancecompared to glioma cell lines U87 and SHG44. Compared to non-GSCs, basal reactive oxygen species formation wasreduced in GSCs, while reactive oxygen species generation was increased in non-GSCs. Following exposure to differentdoses of X-rays under normoxic or hypoxic conditions, GSCs with MTA3 overexpression exhibited greaterradiosensitivity than those with stable integration. Additionally, MTA3 overexpression slightly increased the oxygenenhancement ratio (OER) in GSCs. MTA3 overexpression reduced the immunoreactivity of CD133 and nestin in bothstem cell lines, and increased immunofluorescence staining of GFAP and neuronal class Ⅲ β-tubulin, with statisticallysignificant differences (P<0.05). Conclusions     MTA3 is downregulated in GSCs. Overexpression of MTA3 reduces theradioresistance and stemness of GSCs both in vi

关 键 词：转移相关蛋白3 活性氧 胶质瘤 耐药 

分 类 号：R73[医药卫生—肿瘤]