Background and Aims: Accumulating studies have demonstrated that long noncoding RNA plays a vital role in cancer progression. A previous study reported that LINC00665 was overexpressed and acted as a key tumor promoter in lung cancer, but the role of LINC00665 in gastric cancer (GC) remained uncharacterized. Thus, this study aimed to explore the mechanism of LINC00665 in GC. Methods: LINC00665 expression was explored using the Cancer Genome Atlas (TCGA), and a meta-analysis was conducted to assess the expression and prognostic value of LINC00665 in GC from Gene Expression Omnibus databases and the TCGA dataset. Real-time polymerase chain reaction (RT-PCR) was then conducted to verify the LINC00665 expression in GC tissues and cell lines. The effects of LINC00665 on cell proliferation, invasion, metastasis, and cell cycle in GC were evaluated using the CCK-8, wound healing, Transwell, and flow cytometry assays. In vitro validation was also performed. Results: LINC00665 overexpression was found in GC, and LINC00665 upregulation was significantly related to poor overall survival and disease-free survival. LINC00665 expression was associated with tumor depth, lymph node metastasis, and TNM stage. Univariate and multivariate analyses proved that LINC00665 could be an independent prognostic biomarker in GC. LINC00665 knockdown subsequently inhibited cell proliferation, invasion, and metastasis in GC cell lines; promoted cell apoptosis; and arrested GC cell lines in the G0/G1 phase. Western blot analysis indicated that LINC00665 silencing inhibited epithelial-mesenchymal transition and decreased the expression levels of TGF-beta 1, Smad2, and alpha-SMA. Conclusion: LINC00665 can be a potential diagnostic and prognostic biomarker for GC patients, and LINC00665 promotes GC cell proliferation, invasion, and metastasis by activating the TGF-beta signal pathway.