Knockdown of circHIPK3 Facilitates Temozolomide Sensitivity in Glioma by Regulating Cellular Behaviors Through miR-524-5p/KIF2A-Mediated PI3K/AKT Pathway
Knockdown of circHIPK3 Facilitates Temozolomide Sensitivity in Glioma by Regulating Cellular Behaviors Through miR-524-5p/KIF2A-Mediated PI3K/AKT Pathway
复制标题
敲低circHIPK3通过miR-524-5p/ kif2a介导的PI3K/AKT通路调节细胞行为，促进替莫唑胺在胶质瘤中的敏感性
DOI：
10.1089/cbr.2020.3575
复制
发表时间：
2020-08-21
影响因子：
3.4
通讯作者：
Cui, Xia
中科院分区：
文献类型：
--
作者：
Yin, Hongqian;Cui, Xia
背景：替莫唑胺（TMZ）耐药是脑胶质瘤临床化疗的严重障碍。Circular RNA homeodomain interacting protein kinase 3（circHIPK 3）可能参与胶质瘤的发生发展，但circHIPK 3在TMZ耐药胶质瘤中的分子机制尚不清楚。采用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑法测定TMZ的半数抑制浓度（IC 50）和细胞增殖。流式细胞术检测细胞凋亡，transwell法检测细胞迁移和侵袭。结果：在TMZ耐药的胶质瘤组织和细胞中，cyclHIPK 3表达上调，而在TMZ耐药的胶质瘤细胞中，cyclHIPK 3表达上调。在TMZ耐药的胶质瘤细胞中，敲低circHIPK 3和抑制驱动蛋白家族成员2A（KIF 2A）都可以促进TMZ敏感性和凋亡，但抑制增殖和转移。CircHIPK 3靶向microRNA-524- 5 p（miR-524- 5 p）和KIF 2A作为miR-524- 5 p的下游靶标发挥作用。miR-524- 5 p的减少通过上调KIF 2A减轻si-circHIPK 3对TMZ耐药胶质瘤细胞的作用。结论：cyclHIPK 3基因的下调可通过miR-524 - 5 p/KIF 2A介导的PI 3 K/AKT信号通路调节胶质瘤细胞的增殖、转移和凋亡，从而提高TMZ敏感性。CircHIPK 3可能成为TMZ耐药胶质瘤诊断和治疗的潜在靶点。
Background: Temozolomide (TMZ) resistance is a serious hindrance in clinical chemotherapy for glioma. Circular RNA homeodomain interacting protein kinase 3 (circHIPK3) can be involved in regulating the progression of glioma, but the molecular mechanism of circHIPK3 in TMZ-resistant-glioma is completely unclear.Materials and Methods: The levels of circRNA, miRNA, and mRNA were examined using quantitative real-time polymerase chain reaction. 3-(4, 5-dimethylthiazol-2-y1)-2, 5-diphenyl tetrazolium bromide assay was used for assessing the half inhibitory concentration (IC50) of TMZ and cell proliferation. Cell apoptosis and metastasis (migration and invasion) were detected by flow cytometry and transwell assay, respectively. Western blot and dual-luciferase reporter assay were performed several times to analyze the expression levels of associated proteins and the targeted relation.Results: The upregulation of circHIPK3 was found in TMZ-resistant glioma tissues and cells. Both circHIPK3 knockdown and kinesin family member 2A (KIF2A) inhibition could facilitate TMZ sensitivity and apoptosis but repress proliferation and metastasis in TMZ-resistant glioma cells. CircHIPK3 targeted microRNA-524-5p (miR-524-5p) and KIF2A functioned as a downstream target of miR-524-5p. Decrease of miR-524-5p relieved the effects of si-circHIPK3 on TMZ-resistant glioma cells by upregulating KIF2A. Downregulation of circHIPK3 refrained the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signal pathway partly through miR-524-5p/KIF2A axis.Conclusions: Knockdown of circHIPK3 promoted TMZ sensitivity in glioma by modulating proliferation, metastasis, and apoptosis through miR-524-5p/KIF2A-mediated PI3K/AKT pathway. CircHIPK3 may be the potential target for the diagnosis and therapy of TMZ-resistant glioma.