Ran Yuge, Han Shuai, Gao Dongxue, Chen Xiaobo, Liu Chan
Department of Radiotherapy, Affiliated Hospital of Hebei University, Baoding, Hebei 071000, P.R. China.
Department of Medicine, Batai Biopharmaceutical Co., Ltd., Beijing 102600, P.R. China.
Exp Ther Med. 2024 Jul 23;28(4):373. doi: 10.3892/etm.2024.12662. eCollection 2024 Oct.
Frizzled family protein 2 (FZD2) is widely associated with tumor development and metastasis. The present study aimed to gain an insight into the role and regulatory mechanism of FZD2 in glioma. The expression level of FZD2 in normal astrocyte and glioma cells was determined by reverse transcription-quantitative PCR and western blotting, and cell transfection was conducted for FZD2 expression knockdown. Malignant behaviors including cell proliferation, migration and invasion, vasculogenic mimicry (VM) and cell stemness were determined using Cell Counting Kit-8, 5-Ethynyl-2'-deoxyuridine (EdU) staining, colony formation, wound healing, Transwell, 3D culturing and sphere formation assays. The expression levels of proteins related to stemness, epithelial-mesenchymal transition (EMT) and Notch/NF-κB signaling were measured by western blotting. Then, the Notch agonist, Jagged-1 (JAG), was adopted for rescue experiments. The results demonstrated that FZD2 was highly expressed in glioma cells. Interference of FZD2 expression suppressed the proliferation of glioma cells, as evidenced by the reduced cell viability and the number of EdU cells and colonies. Meanwhile, the reduced sphere formation ability and decreased protein expression of Nanog, Sox2 and Oct4 following FZD2 knockdown confirmed that FZD2 repressed cell stemness in glioma. Additionally, FZD2 knockdown suppressed the migration, invasion, EMT and VM formation capabilities of glioma cells, and also blocked the Notch/NF-κB signaling pathway. Furthermore, activation of Notch by JAG treatment partially reversed the aforementioned FZD2 knockdown-mediated changes in glioma cell malignant behaviors. In conclusion, FZD2 may contribute to glioma progression through activating the Notch/NF-κB signaling pathway, providing a plausible therapeutic target for the treatment of glioma.
卷曲蛋白家族蛋白2（FZD2）与肿瘤的发生和转移密切相关。本研究旨在深入了解FZD2在胶质瘤中的作用及调控机制。通过逆转录定量PCR和蛋白质印迹法检测正常星形胶质细胞和胶质瘤细胞中FZD2的表达水平，并进行细胞转染以敲低FZD2的表达。使用细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷（EdU）染色、集落形成、伤口愈合、Transwell、三维培养和球体形成试验来检测包括细胞增殖、迁移和侵袭、血管生成拟态（VM）和细胞干性在内的恶性行为。通过蛋白质印迹法检测与干性、上皮-间质转化（EMT）和Notch/NF-κB信号相关的蛋白质表达水平。然后，采用Notch激动剂Jagged-1（JAG）进行挽救实验。结果表明，FZD2在胶质瘤细胞中高表达。FZD2表达的干扰抑制了胶质瘤细胞的增殖，细胞活力降低以及EdU阳性细胞和集落数量减少证明了这一点。同时，FZD2敲低后球体形成能力降低以及Nanog、Sox2和Oct4蛋白表达下降证实FZD2抑制了胶质瘤细胞的干性。此外，FZD2敲低抑制了胶质瘤细胞的迁移、侵袭、EMT和VM形成能力，还阻断了Notch/NF-κB信号通路。此外，JAG处理激活Notch部分逆转了上述FZD2敲低介导的胶质瘤细胞恶性行为变化。总之，FZD2可能通过激活Notch/NF-κB信号通路促进胶质瘤进展，为胶质瘤治疗提供了一个合理的治疗靶点。
