Solute carrier (SLC) genes are known to be involved in the development of human tumors. Here, we identified a group of non-small cell lung cancer (NSCLC) - associated SLC genes and explored the role and potential mechanism of action for SLC14A1 in NSCLC.We used public NSCLC gene expression databases to select NSCLC-associated SLC genes. A Cox proportional risk model was used to estimate the hazard rate of selected NSCLC-associated SLC genes. SLC14A1 was selected as a candidate for further study. The expression and effect on survival of SLC14A1 in multiple NSCLC datasets were investigated. The expression of SLC14A1 was modulated in Calu-6 and A549 cells by transfection and a series of proliferation and migration assays were performed. Parameters related to glycolysis and HK2 expression were detected after transfection.Fourteen SLC genes were identified as NSCLC-related SLC genes. Among them, SLC14A1 was downregulated in tumor tissues in multiple datasets. Low expression of SLC14A1 was indicative of poor prognosis in NSCLC. Knockdown of SLC14A1 promoted proliferation and migration in Calu-6 cells. Overexpression of SLC14A1 showed the opposite effect in A549 cells. Additionally, SLC14A1 changes were associated with glycolysis. This was demonstrated by HK2 mRNA and protein levels being regulated by SLC14A1.In our study SLC14A1 suppressed the proliferation and migration of NSCLC cell lines Calu-6 and A549 and was associated with glycolysis. Therefore, SLC14A1 may be a diagnostic marker or therapeutic target for NSCLC patients.© 2022 by the Association of Clinical Scientists, Inc.