AG-1031 induced autophagic cell death and apoptosis in C6 glioma cells associated with Notch-1 signaling pathway
AG-1031 induced autophagic cell death and apoptosis in C6 glioma cells associated with Notch-1 signaling pathway
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AG-1031 诱导与 Notch-1 信号通路相关的 C6 胶质瘤细胞自噬细胞死亡和凋亡
DOI：
10.1002/jcb.26781
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发表时间：
2018
影响因子：
4
通讯作者：
Yang Zhuo
中科院分区：
文献类型：
--
作者：
Wang Yan;Wang Hui;Ge Hui;Yang Zhuo
胶质瘤是最常见的原发性脑肿瘤，发病率高，预后差。AG-1031是一种抗肿瘤药物，其对C6胶质瘤细胞的作用尚不清楚。本研究旨在探讨AG-1031对C6细胞的作用，并试图找出其对C6细胞毒性的潜在机制。3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑（MTT）试验表明，AG-1031以浓度依赖性方式抑制细胞活力，而3-甲基腺嘌呤（3-MA）降低了这种作用。Hoechst 33342染色和Western blot分析结果表明，AG-1031诱导C6细胞凋亡。Western blot结果显示，AG-1031可显著提高LC 3-II/LC 3-I比值，降低P62表达。此外，我们的结果显示，巴弗洛霉素A1增加了AG-1031处理的细胞中LC 3-II的表达，这表明AG-1031可以增加C6细胞中的自噬。同时，Western blot检测AG-1031对Notch-1信号通路的抑制作用。免疫荧光检测显示，Notch胞内区（NICD）的表达也明显降低。此外，Notch-1信号的激活减轻了AG-1031诱导的自噬细胞死亡和凋亡。此外，磷酸化Akt及其下游效应机制靶点雷帕霉素（mTOR）被AG-1031减少。这些结果表明，AG-1031可能通过下调Notch-1信号通路抑制Akt-mTOR信号通路，并通过Notch-1信号通路激活C6细胞凋亡，从而诱导自噬性细胞死亡，这为未来治疗胶质瘤开辟了新的靶点。
Glioma is the most common primary brain tumor with high morbidity and poor prognosis. The effect of AG‐1031, which is developed as an antineoplastic drug, on C6 glioma cells is still not clear. The aim of this research was to explore the effect of AG‐1031 on C6 cells, and tried to find out its potential mechanism on cytotoxicity of C6 cells. The 3‐(4,5‐dimethylthiazol −2‐yl) −2,5‐ diphenyltetrazolium bromide (MTT) assay showed that AG‐1031 inhibited cell viability in a concentration‐dependent manner, whereas 3‐methyadenine (3‐MA) reduced this effect. Results from hoechst 33342 staining and Western blot assay indicated that AG‐1031 induced C6 cell apoptosis. Western blot assay presented that AG‐1031 notably increased the LC3‐II/LC3‐I ratio and decreased the expression of P62. Besides, our results showed that bafilomycin A1 increased the expression of LC3‐II in cells treated with AG‐1031, which indicated that AG‐1031 can increase autophagy in C6 cells. Meanwhile, Western blot assay showed that AG‐1031 can inhibit Notch‐1 signaling by testing relative protein expressions. The expression of the intracellular domain of Notch (NICD) also decreased according to immunofluorescence assay. Additionally, the activation of Notch‐1 signaling alleviated AG‐1031‐induced autophagic cell death and apoptosis. Furthermore, phosphorylated Akt and its downstream effector mechanistic target of rapamycin (mTOR) were reduced with AG‐1031. These results suggest that AG‐1031 may induce autophagic cell death through the inhibition of Akt‐mTOR signaling pathway by down‐regulating Notch‐1 signaling pathway and activating apoptosis in C6 cells via Notch‐1 signaling, which develops a new target spot to treat glioma in the future.