Department of Breast and Thyroid Surgery, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, 301 Yanchangzhong Road, Shanghai, 200072, People's Republic of China.
Cell Mol Biol Lett. 2023 Apr 26;28(1):34. doi: 10.1186/s11658-023-00448-7.
Breast cancer (BC) is a common threat to women. The continuous activation of nuclear factor kappa B (NF-κB) signaling pathway contributes to the development of BC. This study aimed to investigate the role of a circular RNA (circRNF10) in BC progression and regulating NF-κB signaling pathway.
Bioinformatics analysis, RT-qPCR, subcellular fractionation, FISH, RNase R treatment, and actinomycin D assay were used to explore the expression and characteristics of circRNF10 in BC. The biological functions of circRNF10 in BC were analyzed by MTT assay, colony formation assay, wound healing assay, and Transwell assay. RNA pulldown and RIP assay were used to identify the interaction between circRNF10 and DEAH (Asp-Glu-Ala-His) box helicase 15 (DHX15). The impact of circRNF10-DHX15 interaction on NF-κB signaling pathway was explored by western blot, IF, and co-IP. Furthermore, dual-luciferase reporter assay, ChIP, and EMSA were performed to assess the effect of NF-κB p65 on DHX15 transcription.
CircRNF10 was downregulated in BC, and lower expression of circRNF10 was related to poor prognosis of patients with BC. CircRNF10 inhibited the proliferation and migration of BC. Mechanically, circRNF10-DHX15 interaction sequestered DHX15 from NF-κB p65, thereby inhibiting the activation of NF-κB signaling pathway. On the other hand, NF-κB p65 enhanced DHX15 transcription by binding to the promoter of DHX15. Altogether, circRNF10 impaired the DHX15-NF-κB p65 positive feedback loop and suppressed the progression of BC.
CircRNF10-DHX15 interaction suppressed the DHX15-NF-κB p65 positive feedback loop, thereby inhibiting BC progression. These findings provide new insights in the continuous activation of NF-κB signaling pathway and raised potential therapeutic approach for BC treatment.
乳腺癌（BC）是女性面临的常见威胁。核因子 kappa B（NF-κB）信号通路的持续激活有助于 BC 的发展。本研究旨在探讨环状 RNA（circRNF10）在 BC 进展中的作用及其对 NF-κB 信号通路的调控作用。
采用生物信息学分析、RT-qPCR、亚细胞分离、FISH、RNase R 处理和放线菌素 D 测定等方法，探讨 circRNF10 在 BC 中的表达和特征。通过 MTT 测定、集落形成测定、划痕愈合测定和 Transwell 测定分析 circRNF10 在 BC 中的生物学功能。采用 RNA 下拉和 RIP 测定鉴定 circRNF10 与 DEAH（Asp-Glu-Ala-His）盒解旋酶 15（DHX15）之间的相互作用。通过 Western blot、IF 和 co-IP 探讨 circRNF10-DHX15 相互作用对 NF-κB 信号通路的影响。此外，还进行了双荧光素酶报告基因测定、ChIP 和 EMSA 实验，以评估 NF-κB p65 对 DHX15 转录的影响。
circRNF10 在 BC 中表达下调，circRNF10 表达水平较低与 BC 患者预后不良有关。circRNF10 抑制 BC 的增殖和迁移。机制上，circRNF10-DHX15 相互作用将 DHX15 从 NF-κB p65 中隔离出来，从而抑制 NF-κB 信号通路的激活。另一方面，NF-κB p65 通过结合 DHX15 启动子增强 DHX15 的转录。总之，circRNF10 破坏了 DHX15-NF-κB p65 正反馈环，抑制了 BC 的进展。
circRNF10-DHX15 相互作用抑制了 DHX15-NF-κB p65 正反馈环，从而抑制了 BC 的进展。这些发现为 NF-κB 信号通路的持续激活提供了新的见解，并为 BC 的治疗提供了潜在的治疗方法。
