This study investigates the influence of long noncoding RNA HOST2 on the biological functions of gastric cancer cells; including proliferation, migration and invasion. Differentially expressed lncRNAs in gastric cancer (GC) were screened by microarray analysis, and HOST2 expression in GC tissues and cell lines was determined by quantitative real-time PCR (qRT-PCR). GC cell proliferation, migration and invasion were detected by CCK-8, wound healing and transwell assays. Western blot investigated expression of epithelial-mesenchymal transition (EMT) related proteins, and association was established between over-expressed HOST2 and the number of patients with lymph node and distant metastasis. HOST2 expression was also positively related to GC cell invasion ability, and although its expression in the p-shHOST2 group was remarkably decreased, it was significantly higher than in the Mock and NC groups. Compared to the Mock and NC groups, the p-shHOST2 group presented significant decreases in proliferation and wound healing rates, and the reverse result was noted in the p-HOST2 group. In addition, the number of p-shHOST2 group invasive cells was remarkably less than in the Mock and NC group, and the opposite result was achieved in the p-HOST2 group. Moreover, p-HOST2 had more significant EMT, but this was suppressed in the p-shHOST2 group. Finally, HOST2 silencing suppressed GC cell proliferation, migration and invasion; and it could therefore be considered as a novel biomarker and therapeutic target in gastric cancer.