ObjectiveTo explore the mechanism by which LINC00636 promotes the proliferation and metastasis of non-small cell lung cancer (NSCLC) by targeting NM23-H1. MethodsBased on the Affymetrix® GeneChip Human Transcriptome Array 2.0 (HTA2.0), differentially expressed long non-coding RNA (lncRNA) in three pairs of NSCLC tissues with and without lymph node metastasis were analyzed, and LINC00636 was selected as the candidate gene. The effects of LINC00636 knockdown and overexpression on the proliferation, apoptosis, migration and subcutaneous tumorigenicity of PC9 cells were analyzed. The dual-luciferase assay was used to analyze the targeting regulation of LINC00636 on NM23-H1. ResultsThe expression of LINC00636 in the serum and tissues of NSCLC patients with lymph node metastasis was higher than that in patients without lymph node metastasis. LINC00636 knockdown could inhibit the invasion and proliferation of NSCLC cells. LINC00636 could target and regulate the expression of NM23-H1, and the expression of LINC00636 and NM23-H1 in NSCLC tissues and cells was negatively correlated. The inhibitory effect of NM23-H1 on the proliferation and invasion of PC9 cells could be partially reversed by LINC00636 overexpression. ConclusionLINC00636 promotes the proliferation and metastasis of NSCLC cells by inhibiting the expression of NM23-H1 and is a potential prognostic marker and therapeutic target for NSCLC.