Department of Radiation Oncology, The First Affiliated Hospital, Guangzhou Medical University, Guangzhou, China.
Department of Neurosurgery, Kunming Children's Hospital, No. 288, Qianxing Road, Kunming, 650030, China.
Neurochem Res. 2021 Dec;46(12):3222-3246. doi: 10.1007/s11064-021-03427-6. Epub 2021 Aug 21.
Since few reports have mentioned the role of FOSL1 in the radiotherapy sensitivity of glioma, this study would dig deep into this aspect. Cancer stem cells (CSCs) isolated by magnetic bead assay were identified by microscopy, qRT-PCR and western blot. The number of apoptotic cells was counted 72 h after X-ray irradiation to evaluate the sensitivity of cancer cells to radiotherapy. The effects of radiotherapy, FOSL1 and miR-27a-5p on basic cell functions were detected by functional experiments. The expressions of FOSL1, apoptosis-related genes and miR-27a-5p were detected by qRT-PCR and western blot as needed. The differential expression of FOSL1 and the effect of miR-27a-5p on survival rate were analyzed using GEPIA and UALCAN, respectively. FOSL1 was found highly expressed in glioma cells and patients. The appearance of spherical cells and high expressions of CSC-related markers indicated the successful isolation of CSC-like cells. The increment of X-ray dose enhanced the sensitivity of cancer cells to radiotherapy. Radiotherapy down-regulated cell viability and the expressions of FOSL1 and Bcl-2, but up-regulated cell apoptosis and the expressions of cleaved caspase-3 and Bax, which could be partially reversed by overexpressed FOSL1 or further enhanced by shFOSL1. MiR-27a-5p was highly expressed in in patients with glioma, which was associated with poor prognosis, while shFOSL1-inhibited miR-27a-5p expression enhanced the sensitivity of glioma stem cells to radiotherapy. In vivo experiments further verified the results obtained from in vitro experiments. Silent FOSL1 strengthened the radiosensitivity of glioma by down-regulating miR-27a-5p.
由于很少有报道提及 FOSL1 在脑胶质瘤放疗敏感性中的作用，本研究将深入探讨这一方面。通过显微镜、qRT-PCR 和 Western blot 鉴定磁珠分离的肿瘤干细胞（CSC）。照射 X 射线 72 小时后计数凋亡细胞数，以评估癌细胞对放疗的敏感性。通过功能实验检测放疗、FOSL1 和 miR-27a-5p 对细胞基本功能的影响。根据需要通过 qRT-PCR 和 Western blot 检测 FOSL1、凋亡相关基因和 miR-27a-5p 的表达。分别使用 GEPIA 和 UALCAN 分析 FOSL1 的差异表达和 miR-27a-5p 对生存率的影响。结果发现 FOSL1 在脑胶质瘤细胞和患者中高表达。出现球体细胞和 CSC 相关标志物的高表达表明成功分离出 CSC 样细胞。增加 X 射线剂量增强了癌细胞对放疗的敏感性。放疗下调细胞活力和 FOSL1 和 Bcl-2 的表达，但上调细胞凋亡和 cleaved caspase-3 和 Bax 的表达，而过表达 FOSL1 可部分逆转，shFOSL1 进一步增强。miR-27a-5p 在脑胶质瘤患者中高表达，与预后不良相关，而 shFOSL1 抑制 miR-27a-5p 表达增强了脑胶质瘤干细胞对放疗的敏感性。体内实验进一步验证了体外实验的结果。沉默 FOSL1 通过下调 miR-27a-5p 增强了脑胶质瘤的放射敏感性。
