Affiliated Tangshan Gongren Hospital, North China University of Science and Technology, Tangshan, China.
College of Life Science, North China University of Science and Technology, Tangshan, China.
Front Immunol. 2023 Jan 18;13:947136. doi: 10.3389/fimmu.2022.947136. eCollection 2022.
plays an important role in the development of colon cancer. This study aims to evaluate the expression of in colon cancer and discover how it is regulated by transcriptional factors and miRNA.
The expression of was explored by TIMER2.0, UALCAN, and Human Protein Atlas (HPA) databases. TRANSFAC and Contra v3 were used to predict the potential binding sites of transcription factors in the promoter. TargetScan and starBase v2.0 were used to predict the potential binding ability of miRNAs to the 3' untranslated region (3'UTR) of . SurvivalMeth was used to explore the differentially methylated sites in the promoter. Western blotting was used to detect the expression of and . Dual-luciferase reporter assay and chromatin immunoprecipitation (ChIP) assay were performed to determine the targeting relationship of , , or miR-27a-3p with . -related genes were explored by constructing a protein-protein interaction (PPI) network and performing pathway analysis by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG).
was highly expressed in colon cancer tissues. The mRNA and protein expression levels of were reduced by si-. mRNA was obviously reduced by inhibitor and increased by activator. protein was also inhibited by miR-27a-3p. Dual-luciferase reporter assays showed that after knocking down or inhibiting , the promoter activity of was decreased by 21% and 70%, respectively; after activating , the promoter activity of increased by 2.3 times. As or binding site was mutated, the transcriptional activity of was significantly decreased. ChIP assay showed that and combined to the promoter of . The luciferase activity of 3'UTR decreased after being co-transfected with miR-27a-3p mimics and increased by miR-27a-3p antagomir. As the miR-27a-3p binding site was mutated, we did not find any significant effect of miR-27a-3p on reporter activity. PPI network assay revealed a set of -related genes, which included , , , and GO and KEGG analyses revealed that the target genes occur more frequently in immune-related pathways.
Our results indicated that is regulated by , , miR-27a-3p, and several immune-related genes, which in turn affects colon cancer.
在结肠癌的发展中发挥重要作用。本研究旨在评估 在结肠癌中的表达，并发现其如何受转录因子和 miRNA 调控。
通过 TIMER2.0、UALCAN 和 Human Protein Atlas (HPA) 数据库探讨 的表达。TRANSFAC 和 Contra v3 用于预测 启动子中转录因子的潜在结合位点。TargetScan 和 starBase v2.0 用于预测 miRNA 与 3'非翻译区 (3'UTR) 的潜在结合能力。SurvivalMeth 用于探索 启动子中的差异甲基化位点。Western blotting 用于检测 的表达。双荧光素酶报告基因检测和染色质免疫沉淀 (ChIP) 实验用于确定 、 或 miR-27a-3p 与 的靶向关系。通过构建蛋白质-蛋白质相互作用 (PPI) 网络并通过基因本体论 (GO) 和京都基因与基因组百科全书 (KEGG) 进行通路分析来探索 -相关基因。
在结肠癌组织中高表达。si- 降低 的 mRNA 和蛋白表达水平。 抑制剂明显降低 mRNA， 激活剂增加 mRNA。miR-27a-3p 也抑制 蛋白。双荧光素酶报告基因检测显示，敲低 或抑制 后， 的启动子活性分别降低 21%和 70%；激活 后， 的启动子活性增加 2.3 倍。当 或 结合位点发生突变时， 的转录活性显著降低。ChIP 实验显示 与 结合到 的启动子上。共转染 miR-27a-3p 模拟物后， 的 3'UTR 荧光素酶活性降低，而 miR-27a-3p 拮抗剂增加。当 miR-27a-3p 结合位点发生突变时，我们没有发现 miR-27a-3p 对报告基因活性有任何显著影响。PPI 网络分析揭示了一组 -相关基因，包括 、 、 、和 。GO 和 KEGG 分析表明，靶基因更频繁地出现在免疫相关通路中。
我们的结果表明， 受 、 、miR-27a-3p 和几个免疫相关基因的调控，进而影响结肠癌。
