Computational protocol: MicroRNA-671-3p inhibits the development of breast cancer: A study based on in vitro experiments, in-house quantitative polymerase chain reaction and bioinformatics analysis

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[…] The predicted target genes of miR-671-3p were determined using the miRWalk 2.0 database (zmf.umm.uni-heidelberg.de/apps/zmf/mirwalk2), which contains 12 online tools, namely Targetscan, RNAhybrid, RNA22, PITA, Pictar2, miRWalk, Microt4, miRNAMap, miRDB, mirbridge, miRanda and miRMap. Targets predicted by >3 algorithms were selected for GO functional annotation and KEGG pathway analyses using the DAVID online tool (david.ncifcrf.gov). Subsequently, the enriched results were visualized with the R package 'ggplot2' (cran.r-project.org/web/packages/ggplot2/index.html). To better understand the molecular mechanism of miR-671-3p in BC, target genes enriched in cancer-associated pathways were uploaded to the STRING database (string-db.org) for PPI network construction. Meanwhile, the regulatory network of the miRNA-gene was constructed and visualized with Cytoscape version 3.4.0 software (). Furthermore, the expression of these target mRNAs was validated using data from TCGA (cancergenome.nih.gov). A heatmap was drawn with the R package 'pheatmap' (cran.r-project.org/web/packages/pheatmap/index.html), and a box-scatter plot was generated with GraphPad Prism 5 software (GraphPad Software, Inc., La Jolla, CA, USA). […]

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