Changes in the miRNA transcriptome in the test samples in comparison with the control samples were determined using the miRNA microarray technique GeneChip miRNA 2.0 Array (Affymetrix), as described by the manufacturer. Data were analyzed using a microarray scanning GeneArray scanner (Agilent Technologies).
Predictive evaluation of the effect of selected miRNAs on mRNAs were significantly differentiated in test samples from control samples was performed using the TargetScan database (http://www.targetscan.org; accessed on February 15, 2022) (29 (link)) and miRanda (http://mirdb.org; accessed on15 February 2022) (30 (link), 31 (link)). According to the miRDB database, “This is an online database for miRNA target prediction and functional annotations. All the targets in miRDB were predicted using a bioinformatics tool, MirTarget, which was developed by analyzing thousands of miRNA-target interactions from high-throughput sequencing experiments. Common features associated with miRNA binding and target downregulation have been identified and are used to predict miRNA targets with machine learning methods. A predicted target with a prediction score of >80 is most likely to be real; however, if the score is below 60, then one needs to exercise caution, and it is recommended to have other supporting evidence as well” (30 (link), 31 (link)).
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