One challenge in miRNA–genes–diseases interaction studies is that it is challenging to find labeled data that indicate a positive or negative relationship between miRNA and genes. The use of one-class classification methods shows a promising path for validating them. We have applied two one-class classification methods, Isolation Forest and One-class SVM, to validate miRNAs interactions with the ERBB2 gene present in breast cancer scenarios using features extracted via sequence-binding. We found that the One-class SVM outperforms the Isolation Forest model, with values of sensitivity of 80.49% and a specificity of 86.49% showing results that are comparable to previous studies. Additionally, we have demonstrated that the use of features extracted from a sequence-based approach (considering miRNA and gene sequence binding characteristics) and one-class models have proven to be a feasible method for validating these genetic molecule interactions.