The present study aimed to investigate the expression levels and clinical significance of the calcineurin B homologous protein 2 (CHP2) in non-small cell lung cancer (NSCLC), and to study its effects on biological characteristics of NSCLC cells. Tumor and adjacent samples were collected from 196 NSCLC patients. Western blot analysis was used to detect the expression levels of the CHP2 in 8 pairs of NSCLC fresh tissues and 4 NSCLC cell lines. Immunohistochemical analysis was used to detect the expression of the CHP2 in 188 additional pairs of NSCLC wax block tissues. The data indicated that the expression levels of the CHP2 in the paraffin and fresh tissues of NSCLC were significantly higher than those of the adjacent tissues. According to the histo-score, univariate and multivariate analysis indicated that a high expression level of CHP2 was an important factor affecting the 5-year survival rate of NSCLC patients. After knocking down the expression of CHP2 in NSCLC cell lines, the proliferative, migratory, and invasive activities of NSCLC-CHP2 cells were decreased which were assessed by Western blotting, Cell Counting Kit-8, and transwell and wound-healing assays. In conclusion, the data demonstrated that CHP2 was highly expressed in NSCLC and that it could promote the development of NSCLC, suggesting its potential application for the therapy of NSCLC.