环状 RNA 谱分析揭示了 hsa_circ_0000375/miR-7706 通路在乳腺癌中的调控作用。
The investigation of circRNA profiling reveals the regulatory role of the hsa_circ_0000375/miR-7706 pathway in breast cancer.
机构信息
Breast Center, the Fourth Hospital of Hebei Medical University, Shijiazhuang, 050017, Hebei, P.R. China.
Research Center, the Fourth Hospital of Hebei Medical University, Shijiazhuang, 050017, Hebei, P.R. China.
出版信息
Mol Biol Rep. 2023 Dec;50(12):9993-10004. doi: 10.1007/s11033-023-08798-3. Epub 2023 Oct 30.
BACKGROUND
Circular RNAs (circRNAs) take an effect on tumorigenesis and progression. However, circRNAs have not been systematically identified in breast cancer (BC) as crucial regulators in multitudinous biological processes. This study is conducted to explore novel circRNAs in BC and the corresponding mechanisms of their action.
METHODS
The circRNA expression profile and RNA-sequencing data about BC were respectively downloaded from public database. Differentially expressed circRNAs, miRNAs, and mRNAs were identified by fold change filtering. The competing endogenous RNAs (ceRNAs) network was established based on the relationship between circular RNAs, miRNAs and mRNAs. GO and KEGG enrichment analysis of the overlapped genes were carried out to predict the potential functions and mechanisms of circRNAs in BC. The CytoHubba plugin in Cytoscape was applied to identify the hub genes from the PPI regulatory network. Kaplan-Meier plotter was used to perform survival analysis of these hub genes further. Real-time PCR was performed to test the expression of circRNA in BC tissues. Cell function studies including transwell analysis and CCK-8 analysis were used to investigate circRNAs' biological functions.
RESULTS
A total of seven circRNAs exhibiting differential expression were identified in this study. After the intersection between the predicted target miRNA and the down-regulated differential miRNAs (DEmiRNAs), circRNA-miRNA interactions involving 3 circRNAs and 4 miRNAs were identified. Venn diagram was utilized to intersect the predicted target genes of the 4 miRNAs and the down-regulated differential genes in BC, and 149 overlapped genes were screened out ulteriorly. Additionally, we built a protein-protein interaction (PPI) network and selected six hub genes. Moreover, the survival data of BC patients suggested that low expression of ADIPOQ, LPL and LEP were significantly correlated with poor prognosis. Results from real-time PCR indicated that hsa_circ_0000375 was significantly down-regulated in breast cancer tissues. Functional in vitro experiments showed that over-expression of hsa_circ_0000375 can restrain proliferation, migration and invasion abilities of breast cancer cells. Further verification indicated that hsa_circ_0000375 exerted its anti-oncogene effect via sponge of miR-7706.
CONCLUSIONS
This study constructed and analyzed a circRNA-associated ceRNA regulatory network and uncovered that hsa_circ_0000375 exerted its anti-oncogene effect via sponge of miR-7706.
背景
环状 RNA（circRNAs）在肿瘤发生和进展中发挥作用。然而，circRNAs 在乳腺癌（BC）中作为众多生物过程中的关键调节因子尚未被系统地识别。本研究旨在探索 BC 中的新型 circRNAs 及其作用机制。
方法
从公共数据库中分别下载了关于 BC 的 circRNA 表达谱和 RNA-seq 数据。通过折叠变化筛选鉴定差异表达的 circRNAs、miRNAs 和 mRNAs。基于 circRNAs、miRNAs 和 mRNAs 之间的关系构建竞争内源性 RNA（ceRNA）网络。对重叠基因进行 GO 和 KEGG 富集分析，预测 circRNAs 在 BC 中的潜在功能和机制。应用 Cytoscape 中的 CytoHubba 插件从 PPI 调控网络中识别出关键基因。进一步通过 Kaplan-Meier plotter 对这些关键基因进行生存分析。通过实时 PCR 检测 BC 组织中 circRNA 的表达。通过 Transwell 分析和 CCK-8 分析等细胞功能研究来研究 circRNAs 的生物学功能。
结果
本研究共鉴定出 7 个差异表达的 circRNA。在预测的靶 miRNA 与下调的差异 miRNA（DEmiRNAs）之间的交集后，鉴定出涉及 3 个 circRNA 和 4 个 miRNAs 的 circRNA-miRNA 相互作用。利用 Venn 图将 4 个 miRNA 的预测靶基因与 BC 中下调的差异基因进行交集，进一步筛选出 149 个重叠基因。此外，我们构建了一个蛋白质-蛋白质相互作用（PPI）网络，并选择了 6 个关键基因。此外，BC 患者的生存数据表明，ADIPOQ、LPL 和 LEP 的低表达与预后不良显著相关。实时 PCR 结果表明，hsa_circ_0000375 在乳腺癌组织中显著下调。体外功能实验表明，hsa_circ_0000375 的过表达可以抑制乳腺癌细胞的增殖、迁移和侵袭能力。进一步验证表明，hsa_circ_0000375 通过海绵吸附 miR-7706 发挥其抑癌作用。
结论
本研究构建并分析了一个 circRNA 相关的 ceRNA 调控网络，揭示了 hsa_circ_0000375 通过海绵吸附 miR-7706 发挥其抑癌作用。
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