MicroRNAs (miRs) were a series of non-coding RNAs, which could suppress the transcription of mRNAs; the deregulations of miR could regulate the cancer cell proliferation and apoptosis. In the present study we try to investigate the biological effect of miR-375 on the gastric cancer cells. RT-PCR result showed that miR-375 expression was significantly decreased in gastric cancer cells compared with normal gastric cells and microRNA mimic transfection assay was used to investigate the biological effect of miR-375 in the gastric cancer cells. MTT results demonstrated that the induction of miR-375 significantly inhibited the gastric cancer cell proliferation and FACS study indicated the cell proliferation inhibition was attributed to the induction of gastric cancer cell apoptosis. Astrocyte elevated gene-1 (AEG-1) was an important oncogene in multiple aspects of cancer development and progression, our further studies indicated that the transfection of miR-375 mimic significantly decreased the expression of AEG-1 gene both in mRNA level and protein level, which suggest miR-375 regulate the gastric cancer cell through the regulation of AEG-1 gene. In conclusion, our findings suggested that miR-375 targeted AEG-1 gene in gastric cancer cell and regulated the gastric cancer cell growth in vitro, which highlight the therapeutic potential of miR-375 in gastric cancer clinical treatment.