Withaferin A triggers G2/M arrest and intrinsic apoptosis in glioblastoma cells via ATF4-ATF3-CHOP axis
Withaferin A triggers G2/M arrest and intrinsic apoptosis in glioblastoma cells via ATF4-ATF3-CHOP axis
复制标题
醉茄素 A 通过 ATF4-ATF3-CHOP 轴触发胶质母细胞瘤细胞的 G2/M 期阻滞和内在凋亡
DOI：
10.1111/cpr.12706
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发表时间：
2019-10-23
影响因子：
8.5
通讯作者：
Du, Guanhua
中科院分区：
文献类型：
--
作者：
Tang, Qin;Ren, Liwen;Du, Guanhua
目的Withaferin A （WA）是一种从苦参中提取的具有显著抗癌作用的生物活性化合物。然而，WA在多形性胶质母细胞瘤（GBM）中的抗癌机制尚不清楚。材料与方法采用细胞活力法和裸鼠移植法评价WA对GBM的影响，流式细胞术检测细胞凋亡和细胞周期。通过RNA-seq分析、Western blotting、免疫荧光染色、qRT-PCR和siRNA基因沉默来确定WA影响的信号通路。结果Withaferin A在体外和体内均能显著抑制GBM的生长，并通过上调Bim和Bad的表达触发GBM细胞的内在凋亡。WA通过激活p53不依赖的p21上调，使CDK1的Thr（161）去磷酸化，使GBM细胞处于细胞周期的G2/M期。p21的敲低通过下调Bad而非Bim的表达来恢复细胞周期进程和细胞活力。我们证明了WA通过atf4 - atf4 - chop轴诱导内质网（ER）应激，启动了GBM细胞的凋亡和G2/M阻滞。结论我们通过atf4 - atf4 - chop轴揭示了WA激活GBM细胞凋亡和G2/M阻滞的新途径。这一发现对于优化基于wa的GBM预防和/或治疗方案具有重要意义。
Objective Withaferin A (WA) is a bioactive compound with a remarkable anti-cancer effect derived from Withania somnifera, commonly known as ashwagandha. However, the anti-cancer mechanisms of WA in glioblastoma multiforme (GBM) are still unclear. Materials and Methods Cell viability assays and xenografted nude mice were used to evaluate the effects of WA, along with flow cytometry to detect apoptosis and cell cycle of GBM. RNA-seq analysis, Western blotting, immunofluorescence staining, qRT-PCR and siRNA gene silencing were carried out to determine the signalling pathways affected by WA. Results Withaferin A significantly inhibited the growth of GBM in vitro and in vivo and triggered the intrinsic apoptosis of GBM cells by up-regulating expression of Bim and Bad. WA arrested GBM cells at the G2/M phase of the cell cycle through dephosphorylating Thr(161) of CDK1 by activating p53-independent p21 up-regulation. Knockdown of p21 restored cell cycle progression and cell viability by down-regulating the expression of Bad rather than Bim. We demonstrated that endoplasmic reticulum (ER) stress induced by WA through the ATF4-ATF3-CHOP axis, initiated apoptosis and G2/M arrest in GBM cells. Conclusion We revealed a novel pathway that elucidated WA activation of apoptosis and G2/M arrest in GBM cells through the ATF4-ATF3-CHOP axis. This discovery is important for optimization of WA-based regimens for prevention and/or treatment of GBM.