Yang Dan, Fan Yaqin, Xie Beibei, Yang Jie
Faculty of Health, Yantai Nanshan University, Yantai, Shandong 265713, P.R. China.
Department of Obstetrics and Gynecology, Yuncheng County People's Hospital, Heze, Shandong 274700, P.R. China.
Oncol Lett. 2021 May;21(5):346. doi: 10.3892/ol.2021.12607. Epub 2021 Mar 3.
Increasing number of studies have suggested that microRNA (miR)-203 is a potential prognostic marker for breast cancer. However, the specific molecular mechanism underlying the effects of miR-203 remains unknown. The present study aimed to explore the molecular target and underlying mechanisms of action of miR-203 in breast cancer via bioinformatics analysis and cellular assays, such as wound healing assay and western blotting. In the present study, 17 candidate target genes of miR-203 were identified in the downregulated differentially expressed genes from Affymetrix microarray and TargetScan 7.2 database. Subsequently, FK506 binding protein 5 (FKBP5) was considered as the miR-203 target by 3 different hub gene analysis methods (EcCentricity, Betweenness and Stress). FKBP5 protein expression was significantly downregulated in SUM159 cells transfected with miR-203 mimics compared with SUM159 cells transfected with miR-203 negative control (NC) in western blot analysis. High expression of FKBP5 was associated with poor prognosis in breast cancer based on the results obtained from the Kaplan-Meier Plotter database. In addition, the wound healing assay indicated that the inhibition of migration due to miR-203 overexpression in SUM159 cells was reversed by FKBP5 overexpression. These results suggested that miR-203 may directly target FKBP5. In addition, Gene Set Enrichment Analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that miR-203 might play a role in breast cancer via the 'fatty acid degradation' KEGG pathway. Notably, the levels of fatty acids were significantly reduced in SUM159 cells transfected with miR-203 mimics compared with SUM159 cells transfected with miR-203 NC when assessed by the fatty acid content assay. Finally, virtual screening analysis revealed that ZINC000003944422 may be a potential inhibitor of FKBP5. In summary, the present study demonstrated that miR-203 may directly target FKBP5 in breast cancer via fatty acid degradation and potential drugs, hence providing a novel treatment approach for breast cancer.
越来越多的研究表明，微小RNA（miR）-203是乳腺癌潜在的预后标志物。然而，miR-203发挥作用的具体分子机制仍不清楚。本研究旨在通过生物信息学分析和细胞实验，如伤口愈合实验和蛋白质印迹法，探索miR-203在乳腺癌中的分子靶点及潜在作用机制。在本研究中，从Affymetrix微阵列和TargetScan 7.2数据库中下调的差异表达基因中鉴定出17个miR-203的候选靶基因。随后，通过3种不同的枢纽基因分析方法（中心性、介数和应力）将FK506结合蛋白5（FKBP5）视为miR-203的靶点。在蛋白质印迹分析中，与转染miR-203阴性对照（NC）的SUM159细胞相比，转染miR-203模拟物的SUM159细胞中FKBP5蛋白表达显著下调。基于Kaplan-Meier Plotter数据库的结果，FKBP5的高表达与乳腺癌的不良预后相关。此外，伤口愈合实验表明，SUM159细胞中miR-203过表达导致的迁移抑制可被FKBP5过表达逆转。这些结果表明，miR-203可能直接靶向FKBP5。此外，基因集富集分析和京都基因与基因组百科全书（KEGG）通路富集分析显示，miR-203可能通过“脂肪酸降解”KEGG通路在乳腺癌中发挥作用。值得注意的是，通过脂肪酸含量测定评估，与转染miR-203 NC的SUM159细胞相比，转染miR-203模拟物的SUM159细胞中脂肪酸水平显著降低。最后，虚拟筛选分析表明ZINC000003944422可能是FKBP5的潜在抑制剂。总之，本研究表明，miR-203在乳腺癌中可能通过脂肪酸降解和潜在药物直接靶向FKBP5，从而为乳腺癌提供了一种新的治疗方法。
