Abstract: Objective To explore the function and mechanism of thioredoxin-related protein 14 kDa （TRP14） in the cells proliferation and migration of non-small cell lung cancer （NSCLC）. Methods Analysis the expression level of TRP14 in NSCLC within The Cancer Genome Atlas （TCGA） data. The expression of TRP14 in lung epithelial cells and NSCLC cells were detected by quantitative polymerase chain reaction （qPCR）. Knockdown the TRP14 gene to detect the proliferation of NSCLC cells H1299 and H1975 by CCK8 assay and clone formation assay, and detect the migration and apoptosis by transwell and flow cytometry. After TRP14 was interfered, the downstream pathway genes were detected by western blot to analyze the possible mechanism of TRP14 gene. Results TRP14 expression was significantly increased in NSCLC tissues compared with adjacent tissues, and TRP14 knockdown inhibited cell proliferation and migration, and promoted cell apoptosis. Moreover, TRP14 may play a role by regulating autophagy. Conclusions TRP14 gene played an important role in the cells proliferation and migration of NSCLC by influencing the mechanism of autophagy. TRP14 gene was a new potential molecular diagnostic index and target for NSCLC.