Objective To screen the differentially expressed microRNAs (miRNAs) in colorectal cancer tissues with high expression of gastrin (GAS) by gene chip technique. Methods The level of GAS in tumor tissues from 71 patients with colorectal cancer was detected by enzyme linked immumosorbent assay (ELISA), and the relationship between positive expression of GAS and clinicopathological features was analyzed. Human colorectal cancer tissues with high positive GAS expression (high-expression group, n=4) or negative GAS expression (control group, n=4) were analyzed by miRNA microarray, and the selected differentially expressed miRNAs were validated by qPCR. Results There were 17 cases (23.9%) with GAS content ≥50.00 pg/g (positive GAS expression), while the remaining 54 cases (76.1%) had negative GAS expression. The expression level of GAS was related to the degree of differentiation, Dukes stage and histological type (P<0.05). There were 236 miRNAs had significant differential expression in the high-expression group (GAS content >200.00 pg/g) compared with the control group, of which 159 miRNAs were up-regulated and 77 were down-regulated. A total of 24 miRNAs were screened out, which differentially expressed more than 3 folds in the high-expression group, with 17 up-regulated and 7 down-regulated. Three up-regulated or down-regulated miRNAs were selected for qPCR verification, and the results were consistent with the microarray analysis. Conclusion The miRNAs are differentially expressed in the colorectal cancer tissues with high expression of GAS, and they would be potential targets for the treatment of the colorectal cancer with high GAS expression.