Purpose <p>To investigate the effects of YKT6 on lung cancer proliferation, migration, invasion, and autophagy.</p> Methods <p>Differential expression analysis, clinical characteristics correlation analysis, Kaplan–Meier survival analysis, and univariate and multivariate Cox regression analyses were performed to explore the expression and clinical significance of YKT6 in lung cancer. The clone formation assay, MTT assay, scratch assay, transwell assay, AO fluorescence staining assay and western blot assay was detect the effect of YKT6 on cell proliferation, migration, invasion and autophagy. YKT6 co-expressed genes were collected by LinkedOmics database and analyzed by GO and KEGG enrichment to explore the role of YKT6 in the regulation of lung cancer pathways. Flow cytometry and western blot experiments were used to detect the cell cycle changes and the expression levels of cell cycle-related proteins in knockdown YKT6 cells.</p> Results <p>YKT6 was highly expressed in lung cancer tissues, and its expression was correlated with clinical characteristics. The high expression of YKT6 led to a poor prognosis of lung cancer patients. YKT6 affected the proliferation, migration, invasion and autophagy of lung cancer cells. YKT6 impacted the disease progression of lung cancer through cell cycle regulation. The high expression of YKT6 increased the expression of Cyclin A protein and Cyclin D1 protein expression levels, while inhibiting the expression of p21 protein and p27 protein.</p> Conclusions <p>YKT6 is highly expressed in lung cancer. YKT6 promotes proliferation, migration and invasion, and inhibits autophagy in lung cancer.</p>