Natural killer (NK) cells play significant roles in spontaneous antitumor response in multiple cancers, including gastric cancer. Currently, lncRNAs were identified as essential modulators in the development of NK cells via competing for the target miRNA. However, the regulatory mechanism of GAS5 in NK cells remains largely elusive. The expressions of GAS5 and miR-18a in NK cells were measured by qRT-PCR. The killing effects of NK cells were conducted by lactate dehydrogenase (LDH) assay. Detection of IFN-γ and TNF-α level was carried out using ELISA assay. The interaction between GAS5 and miR-18a was determined by the luciferase reporter system and RIP assay, respectively. We found that GAS5 expression was downregulated while miR-18a expression was upregulated in primary NK cells isolated from GC patient compared with the healthy controls. Moreover, activation of NK cells stimulated by IL-2 enhanced the secretion of IFN-γ, TNF-α, and the expression of GAS5. The deficiency of GAS5 significantly suppressed the secretion of IFN-γ and TNF-α as well as the killing effect of NK cells. Subsequently, luciferase reporter and RIP assay confirmed the interaction between GAS5 and miR-18a. In addition, miR-18a inhibitor attenuated GAS5 silencing induced inhibition of the cytotoxicity of activated NK cells. In conclusion, GAS5 promotes the killing effect of the natural killer cells against GC by regulating miR-18a, providing promising strategies for NK cells based antitumor therapies.