MicroRNAs (miRNAs) have been shown to regulate many cellular processes and to play a role in host-pathogen interactions. However, the role of miRNA in urinary tract infection (UTI) remains unclear. The aim of this study was to analyze and compare miRNAs from supernatants of human bladder epithelial cells infected with ESBL-producing (ESBL019) and non-ESBL-producing (CFT073) uropathogenic E. coli (UPEC) strains and to identify miRNA target genes in human cells and uropathogenic bacteria. In total, 402 unique miRNAs were found. The statistical analysis showed differential expression of 30 miRNAs from bladder cells stimulated with ESBL019, while stimulation with CFT073 did not show any significantly expressed miRNAs when compared to unstimulated controls. The 30 differentially expressed miRNAs in ESBL019 stimulated cells showed 747 predicted individual human gene targets. KEGG and REACTOME pathways showed enrichments in pathways mainly connected to immune regulation and stress responses. Of the 30 differentially expressed host miRNAs, nine miRNAs were found to interact with predictive targets of the whole genome from the multi-resistant, ESBL-producing UPEC strain EC958. This study shows that ESBL019-infected bladder epithelial cells release miRNAs with predictive targets in both human and bacterial genes, although their role in UTI cross-species interactions remains to be clarified.