Inhibition of Ciliogenesis Enhances the Cellular Sensitivity to Temozolomide and Ionizing Radiation in Human Glioblastoma Cells
Inhibition of Ciliogenesis Enhances the Cellular Sensitivity to Temozolomide and Ionizing Radiation in Human Glioblastoma Cells
复制标题
抑制纤毛发生可增强人胶质母细胞瘤细胞对替莫唑胺和电离辐射的敏感性
DOI：
10.3967/bes2022.058
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发表时间：
2022
影响因子：
3.5
通讯作者：
He Jinpeng
中科院分区：
文献类型：
--
作者：
Wei Li;Ma Wei;Cai Hui;Peng Shaopeng;Tian Huanbing;Wang Jufang;Gao Lan;He Jinpeng
目的探讨原发纤毛对胶质母细胞瘤(GBM)细胞对替莫唑胺(TMZ)和电离辐射(IR)反应的调节作用。方法用TMZ或X射线/碳离子处理GBM细胞。原代纤毛用Ar13b和γ-微管蛋白免疫组织化学染色，细胞存活率或存活率测定细胞抵抗能力。结合IFT88消融、水合氯醛和氯化锂诱导的纤毛消融，用吖啶橙染色法检测自噬。用γH_2AX斑点动力学曲线评价细胞的DNA损伤修复能力，用免疫组织化学方法检测DNA依赖蛋白激酶(DNA-PK)的活性。结果原生纤毛在基底膜中保存较多，诱导纤毛生成抑制细胞增殖。TMZ和IR以剂量和时间依赖的方式促进纤毛发生，抑制纤毛发生显著增强细胞对TMZ和IR的敏感性。纤毛发生的抑制通过损害自噬和DNA损伤修复增加了TMZ和IR的致死效应。纤毛发生的干扰降低了DNA-PK的活性，DNA-PK的敲除导致纤毛的形成和伸长。结论原代纤毛通过介导自噬和DNA损伤修复，在调节细胞对TMZ和IR的敏感性中发挥重要作用。
Objective To investigate the function of primary cilia in regulating the cellular response to temozolomide (TMZ) and ionizing radiation (IR) in glioblastoma (GBM). Methods GBM cells were treated with TMZ or X-ray/carbon ion. The primary cilia were examined by immunostaining with Arl13b and γ-tubulin, and the cellular resistance ability was measured by cell viability assay or survival fraction assay. Combining with cilia ablation by IFT88 depletion or chloral hydrate and induction by lithium chloride, the autophagy was measured by acridine orange staining assay. The DNA damage repair ability was estimated by the kinetic curve of γH2AX foci, and the DNAdependent protein kinase (DNA-PK) activation was detected by immunostaining assay. Results Primary cilia were frequently preserved in GBM, and the induction of ciliogenesis decreased cell proliferation. TMZ and IR promoted ciliogenesis in dose- and time-dependent manners, and the suppression of ciliogenesis significantly enhanced the cellular sensitivity to TMZ and IR. The inhibition of ciliogenesis elevated the lethal effects of TMZ and IR via the impairment of autophagy and DNA damage repair. The interference of ciliogenesis reduced DNA-PK activation, and the knockdown of DNA-PK led to cilium formation and elongation. Conclusion Primary cilia play a vital role in regulating the cellular sensitivity to TMZ and IR in GBM cells through mediating autophagy and DNA damage repair.