Circ_0000518 regulates miR-326/EPHB3 axis to promote colorectal cancer progression

IF 1 Q4 GENETICS & HEREDITY

Gene Reports Pub Date : 2024-06-20 DOI:10.1016/j.genrep.2024.101960

Qiang Pang, Jiaqing Cao

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引用次数: 0

Abstract

Background

Circular RNAs (circRNAs) have emerged as potential clinical targets in cancer therapy, since they have been shown to play a crucial role in tumor development by acting as competitive endogenous RNAs (ceRNAs) that regulate downstream mRNA expression by sequestering microRNAs (miRNAs). The present study, aimed to identify novel ceRNA networks involving circRNAs, miRNAs, and mRNAs in using a combination of Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) datasets and explore the function of circ_0000518 in the malignancy of colorectal cancer (CRC).

Methods

Raw data related to CRC, including circRNAs, miRNAs and mRNAs, were downloaded from GEO and TCGA databases. A circRNA/miRNA/mRNA network was established by differential analysis and multiple databases were used to predict downstream targets. The role of this network in the development of CRC was further validated by a series of cellular experiments including dual luciferase assay, migration assay and flow apoptosis assay.

Results

Differential expression analysis of two GEO datasets (GSE142837 and GSE126094) revealed four circRNAs, which were used to predict downstream miRNAs and mRNAs based on several databases. These predictions, combined with TCGA analysis, were used to construct a ceRNA network composed of 4 circRNAs, 9 miRNAs, and 52 mRNAs. The circ_0000518/miR-326/ephrin type-B receptor 3 (EPHB3) axis was eventually selected for functional validation in two CRC cell lines. Subsequently, it was experimentally verified that circ_0000518 was highly expressed in CRC; meanwhile, it was found that the proliferation and migration of CRC cells were significantly weakened by knocking down circ_0000518, while apoptosis of tumors was promoted.

Conclusion

The data demonstrated the identification of a novel circRNA/miRNA/mRNA competitive endogenous network in CRC. The regulatory role of the circ_0000518/miR-326/EPHB3 axis in this network was preliminarily validated by cellular experiments exploring CRC tissue samples and two CRC cell lines.

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Circ_0000518调控miR-326/EPHB3轴，促进结直肠癌进展

背景环状RNA（circRNA）已成为癌症治疗的潜在临床靶点，因为它们作为竞争性内源性RNA（ceRNA），通过封存microRNA（miRNA）调控下游mRNA的表达，在肿瘤发生发展过程中发挥着至关重要的作用。本研究旨在结合基因表达总库（GEO）和癌症基因组图谱（TCGA）数据集，鉴定涉及circRNAs、miRNAs和mRNAs的新型ceRNA网络，并探讨circ_0000518在结直肠癌（CRC）恶性肿瘤中的功能。方法从GEO和TCGA数据库下载与CRC相关的原始数据，包括circRNAs、miRNAs和mRNAs。方法从 GEO 和 TCGA 数据库中下载了与 CRC 相关的原始数据，包括 circRNA、miRNA 和 mRNA，通过差异分析建立了 circRNA/miRNA/mRNA 网络，并利用多个数据库预测下游靶点。结果对两个 GEO 数据集（GSE142837 和 GSE126094）进行差异表达分析，发现了四个 circRNA，并根据多个数据库预测了其下游的 miRNA 和 mRNA。这些预测与 TCGA 分析相结合，构建了一个由 4 个 circRNA、9 个 miRNA 和 52 个 mRNA 组成的 ceRNA 网络。最终选择了 circ_0000518/miR-326/ephrin type-B receptor 3 (EPHB3) 轴在两个 CRC 细胞系中进行功能验证。随后，实验验证了circ_0000518在CRC中的高表达，同时发现通过敲除circ_0000518，CRC细胞的增殖和迁移明显减弱，而肿瘤细胞的凋亡得到促进。通过对 CRC 组织样本和两种 CRC 细胞系进行细胞实验，初步验证了 circ_0000518/miR-326/EPHB3 轴在该网络中的调控作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Gene Reports Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

3.30

自引率

7.70%

发文量

246

审稿时长

49 days

期刊介绍： Gene Reports publishes papers that focus on the regulation, expression, function and evolution of genes in all biological contexts, including all prokaryotic and eukaryotic organisms, as well as viruses. Gene Reports strives to be a very diverse journal and topics in all fields will be considered for publication. Although not limited to the following, some general topics include: DNA Organization, Replication & Evolution -Focus on genomic DNA (chromosomal organization, comparative genomics, DNA replication, DNA repair, mobile DNA, mitochondrial DNA, chloroplast DNA). Expression & Function - Focus on functional RNAs (microRNAs, tRNAs, rRNAs, mRNA splicing, alternative polyadenylation) Regulation - Focus on processes that mediate gene-read out (epigenetics, chromatin, histone code, transcription, translation, protein degradation). Cell Signaling - Focus on mechanisms that control information flow into the nucleus to control gene expression (kinase and phosphatase pathways controlled by extra-cellular ligands, Wnt, Notch, TGFbeta/BMPs, FGFs, IGFs etc.) Profiling of gene expression and genetic variation - Focus on high throughput approaches (e.g., DeepSeq, ChIP-Seq, Affymetrix microarrays, proteomics) that define gene regulatory circuitry, molecular pathways and protein/protein networks. Genetics - Focus on development in model organisms (e.g., mouse, frog, fruit fly, worm), human genetic variation, population genetics, as well as agricultural and veterinary genetics. Molecular Pathology & Regenerative Medicine - Focus on the deregulation of molecular processes in human diseases and mechanisms supporting regeneration of tissues through pluripotent or multipotent stem cells.