Objective To summarize the stemness regulation mechanism of microRNA on invasion, metastasis and chemoresistance of gastric cancer stem cells (GCSCs), and to explore the anti-tumor therapy based on miRNA targeting GCSCs. Method The literatures about the research progress of miRNA and GCSCs at home and abroad in recent years were collected and reviewed. Results MiRNA could regulate a series of important cellular processes such as proliferation, apoptosis, differentiation and epithelial-mesenchymal transition of GCSCs by participating in the expression of related target genes, which was associated with poor prognosis and high mortality of gastric cancer patients. Silencing or restoring the expression of candidate miRNA of GCSCs could provide a novel and promising approach for the treatment of gastric cancer. Conclusions GCSCs have an important relationship with the malignant biological behavior of gastric cancer, and studies have confirmed that miRNA play an important regulatory role in GCSCs. Therefore, miRNA can be used as a potential target for the treatment of gastric cancer. By regulating the expression of specific miRNA, it can inhibit tumor invasion and metastasis, and improve the sensitivity of chemotherapy drugs.