General Clinical Research Center, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.
Department of Laboratory Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.
Cell Death Dis. 2024 Feb 20;15(2):153. doi: 10.1038/s41419-024-06527-7.
Breast cancer (BC) is the most commonly diagnosed malignant tumour in females worldwide. Although remarkable advances in early detection and treatment strategies have led to decreased mortality, recurrence and metastasis remain the major causes of cancer death in BC patients. Increasing evidence has demonstrated that circular RNAs (circRNAs) play critical roles in cancer progression. However, the detailed biological functions and molecular mechanisms of circRNAs in BC are unclear. The aim of this study was to investigate the possible role of circRNAs in the progression of BC. Differentially expressed circRNAs in BC were identified by integrating breast tumour-associated somatic CNV data and circRNA high-throughput sequencing. Aberrant hsa_circ_0007990 expression and host gene copy number were detected in BC cell lines via quantitative polymerase chain reaction (qPCR). The expression level of hsa_circ_0007990 in BC tissues was validated by in situ hybridization (ISH). Loss- and gain-of-function experiments were performed in vitro and in vivo, respectively, to explore the potential biological function of hsa_circ_0007990 in BC. The underlying mechanisms of hsa_circ_0007990 were investigated through MS2 RNA pull-down, RNA immunoprecipitation, RNA fluorescence in situ hybridization, immunofluorescence, chromatin immunoprecipitation and luciferase reporter assays. The levels of hsa_circ_0007990 were elevated in BC tissues and cell lines, an effect that was partly due to host gene copy number gains. Functional assays showed that hsa_circ_0007990 promoted BC cell growth. Mechanistically, hsa_circ_0007990 could bind to YBX1 and inhibit its degradation by preventing ubiquitin/proteasome-dependent degradation, thus enhancing the expression of the cell cycle-associated gene E2F1. Rescue experiments suggested that hsa_circ_0007990 promoted BC progression through YBX1. In general, our study demonstrated that hsa_circ_0007990 modulates the ubiquitination and degradation of YBX1 protein and further regulates E2F1 expression to promote BC progression. We explored the possible function and molecular mechanism of hsa_circ_0007990 in BC and identified a novel candidate target for the treatment of BC.
乳腺癌(BC)是全球女性中最常见的恶性肿瘤。尽管在早期检测和治疗策略方面取得了显著进展,降低了死亡率,但复发和转移仍然是 BC 患者癌症死亡的主要原因。越来越多的证据表明,环状 RNA(circRNA)在癌症进展中发挥着关键作用。然而,circRNA 在 BC 中的详细生物学功能和分子机制尚不清楚。本研究旨在探讨 circRNA 在 BC 进展中的可能作用。通过整合乳腺肿瘤相关体细胞 CNV 数据和 circRNA 高通量测序,鉴定出 BC 中的差异表达 circRNA。通过实时定量聚合酶链反应(qPCR)检测 BC 细胞系中 hsa_circ_0007990 的表达和宿主基因拷贝数。通过原位杂交(ISH)验证 BC 组织中 hsa_circ_0007990 的表达水平。分别在体外和体内进行缺失和获得功能实验,以探讨 hsa_circ_0007990 在 BC 中的潜在生物学功能。通过 MS2 RNA 下拉、RNA 免疫沉淀、RNA 荧光原位杂交、免疫荧光、染色质免疫沉淀和荧光素酶报告基因检测等方法研究了 hsa_circ_0007990 的潜在机制。BC 组织和细胞系中 hsa_circ_0007990 的水平升高,部分原因是宿主基因拷贝数增加。功能测定表明,hsa_circ_0007990 促进 BC 细胞生长。在机制上,hsa_circ_0007990 可以与 YBX1 结合,并通过阻止泛素/蛋白酶体依赖性降解来抑制其降解,从而增强细胞周期相关基因 E2F1 的表达。挽救实验表明,hsa_circ_0007990 通过 YBX1 促进 BC 进展。总的来说,我们的研究表明,hsa_circ_0007990 调节 YBX1 蛋白的泛素化和降解,进一步调节 E2F1 的表达,从而促进 BC 的进展。我们探讨了 hsa_circ_0007990 在 BC 中的可能功能和分子机制,并确定了一个新的治疗 BC 的候选靶点。