NMDA receptor signaling induces the chemoresistance of temozolomide via upregulation of MGMT expression in glioblastoma cells
NMDA receptor signaling induces the chemoresistance of temozolomide via upregulation of MGMT expression in glioblastoma cells
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DOI：
10.1007/s11060-022-04154-w
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发表时间：
2022-10
影响因子：
3.9
通讯作者：
Shohei Tsuji;Shinsuke Nakamura;K. Shoda;Tetsuya Yamada;M. Shimazawa;N. Nakayama;T. Iwama;H. Hara
中科院分区：
文献类型：
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作者：
Shohei Tsuji;Shinsuke Nakamura;K. Shoda;Tetsuya Yamada;M. Shimazawa;N. Nakayama;T. Iwama;H. Hara
目的烷化剂替莫唑胺（TMZ）对胶质母细胞瘤（GBM）患者的预后有显著影响。因此，最大化TMZ功效对于GBM治疗是重要的。许多报道表明，谷氨酸信号通过谷氨酸受体（包括N-甲基-D-天冬氨酸受体（NMDAR））促进GBM进展。虽然NMDAR促进GBM细胞的细胞迁移和侵袭，但它们在TMZ抗性中的作用仍不清楚。因此，我们专注于NMDAR信号和研究其对TMZ resistance.MethodsWe的影响，研究NMDAR信号对O 6-甲基鸟嘌呤DNA甲基转移酶（MGMT），DNA修复酶，诱导化疗耐药性TMZ，使用定量实时聚合酶链反应和蛋白质印迹法在人GBM T98 G细胞。此外，我们使用美金刚胺（MEM），一个NMDAR拮抗剂，研究TMZ/MEM combination.ResultsActivation的NMDAR由N-甲基-D-天冬氨酸（NMDA）的细胞毒性效应的MGMT的表达升高，抑制TMZ在T98 G细胞的效果。相反，通过NMDAR 1 shRNA敲低NMDAR可降低T98 G细胞中MGMT的表达，并增强TMZ的作用。MEM可增强TMZ对T98 G细胞的杀伤作用。MEM抑制NMDAR可降低TMZ诱导的MGMT表达，增加TMZ诱导的DNA烷基化作用。结论NMDAR信号通过上调TMZ诱导的GBM细胞MGMT表达而诱导TMZ耐药。此外，MEM抑制TMZ诱导的MGMT上调，并增加TMZ对MGMT阳性细胞的细胞毒性作用。本研究表明，TMZ和MEM的组合可能是一种新的治疗策略，MGMT阳性GBM。NMDAR信号传导控制MGMT的表达和TMZ的细胞毒性作用。
PurposeThe alkylating agent temozolomide (TMZ) has a significant impact on the prognosis of glioblastoma (GBM) patients. Therefore, maximizing TMZ efficacy is important for GBM treatment. Many reports have shown that glutamate signaling promotes GBM progression via glutamate receptors, including N-methyl-D-aspartate receptors (NMDARs). Although NMDARs promote cell migration and invasion of GBM cells, their role in TMZ resistance remains unclear. Therefore, we focused on NMDAR signaling and investigated its effects on TMZ resistance.MethodsWe investigated the effect of NMDAR signaling on O6-methylguanine DNA methyltransferase (MGMT), a DNA repair enzyme that induces chemoresistance to TMZ, using quantitative real-time polymerase chain reaction and western blotting in human GBM T98G cells. In addition, we used memantine (MEM), an NMDAR antagonist, to investigate the cytotoxic effect of TMZ/MEM combination and its detailed mechanism.ResultsActivation of NMDAR byN-methyl-D-aspartate (NMDA) elevated MGMT expression and suppressed the effect of TMZ in T98G cells. In contrast, knockdown of NMDAR by NMDAR1 shRNA decreased MGMT expression and enhanced the effect of TMZ in T98G cells. The cytotoxic effect of TMZ was enhanced by MEM in T98G cells. Inhibition of NMDAR by MEM decreased MGMT expression and increased DNA alkylation by TMZ.ConclusionNMDAR signaling induced chemoresistance of TMZ via the upregulation of MGMT expression in GBM cells. Furthermore, MEM inhibited TMZ-induced MGMT upregulation and increased the cytotoxic effect of TMZ on MGMT-positive cells. This study demonstrates that the combination of TMZ and MEM could be a new therapeutic strategy for MGMT-positive GBM.Graphical abstractOverview of this study. NMDAR signaling controls the expression of MGMT and the cytotoxic effect of TMZ.