Objective To analyse the expression profile of specific microRNA (miRNA) of gastric cancer tissues. Methods miRNA microarrays were employed to detect the expression profile of miRNA of 21 paired gastric cancer tissues and corresponding paracancerous normal tissues, and preliminary bioinformatic analyses were conducted. The microarray results were validated by Real-Time PCR. Results It was revealed by miRNA microarray detection that there were 88 miRNA with differential expression in gastric cancer tissues and paracancerous normal tissues, among which miR-21, miR-196b, miR-301a, miR-431*, miR-550*, miR-18a and miR-135b were significantly up-regulated, and miR-139-3p, miR-628-3p, miR-596, miR-99b* and miR-638 were significantly down-regulated. The findings of RT-PCR on 4 miRNA (miR-181 and miR-19b of up-regulation and miR-134 and miR-31 of down-regulation) were in high concordance with microarray results. Conclusion There is specific miRNA expression profile of gastric cancer tissues, and these differentially expressed miRNA may serve as new biomarkers of gastric cancer.