The present study aimed to identify microRNA (miRNA) expression profiles associated with multidrug resistance (MDR) in gastric carcinoma. A 5‑fluorouracil (5‑Fu)‑induced MDR gastric cancer cell line was selected and miRNA expression profiling of the cell line was conducted following exposure to 5‑Fu. The miRNA expression profiles between the parental and resistant gastric cancer cells were analyzed by Human miRNA OneArray® v3 and the results were confirmed by quantitative real‑time RT‑PCR. The expression of 9 miRNAs (miR‑10b, ‑22, ‑31, ‑133b, ‑190, ‑501, ‑615, ‑501‑5p and ‑615‑5p) was upregulated while the expression of 18 additional miRNAs (miR‑32, ‑197, ‑210, ‑766, ‑1229, ‑1238, ‑3131, ‑3149, ‑1224‑3p, ‑3162‑3p, ‑532, ‑877, ‑4701‑5p, ‑5096, ‑4728‑3p, ‑1273d, ‑486‑3p and‑4763‑3p) was downregulated in the SGC‑7901/5‑Fu cell line compared with its parental cell line. The results indicate that miRNA expression correlates with MDR in gastric cancer and may serve as biomolecular targets for MDR elimination.