Docosahexaenoic acid (DHA) and linoleic acid (LA) have modulatory effects on breast cancer (BC) cell lines. We aimed to investigate the effects of DHA, LA alone, in combination, and in the presence of paclitaxel on the expression of five microRNAs involved in the pathology of BC in MDA-MB-231 cell line.MDA-MB-231 cells were treated with either DHA or LA or in combination in the presence/absence of paclitaxel (Taxol). Total RNA was extracted and cDNA synthesized from the cells before and after treatment. The expression levels of miR-30, miR-106b, miR-20, miR-126, and miR-194 were determined by quantitative real-time PCR (qPCR).Treatment of MDA-MB-231 cells with DHA modulated the gene expression of miR-30 (increased by 7.74-fold (p 0.0001), miR-194 (decreased by 11-fold (p 0.0001)), miR-106b (increased by 2.64-fold (p = 0.0004), miR-126 (decreased by 50-fold (p 0.0001)), and miR-20 (decreased by 4-fold (p 0.0001)). Additionally, treatment of MDA-MB-231 cells with LA modulated the gene expression of miR-30 (increased by 2.38-fold (p = 0.0001)), miR-194 (decreased by 100-fold (p 0.0001)), miR-106b (decreased by 10-fold (p 0.0001)). The combined DHA/LA treatment of MDA-MB-231 cells showed regulatory effect on the expression of studied microRNAs in which decreased the expression of miR-30 (5.5-fold (p 0.0001)), miR-194 (11-fold (p 0.0001)), miR-20 (3.5-fold (p = 0.0006)), and increased the expression of miR-106b (9.78-fold (p 0.0001)).Modulation of the expression levels of BC-involved microRNAs could be one of the possible mechanisms of action through which DHA and LA may exert their biologic effects on MDA-MB-231 cell line.