The aim of the research was to investigate the expression of lncRNA EPB41L4A-AS2 in non-small cell lung cancer (NSCLC) and evaluate its influence on the proliferation, invasion and apoptosis of NSCLC. A total of 56 NSCLC tissues and its corresponding adjacent tissues were collected. Quantitative Reverse transcription polymerase chain reaction (qRT-PCR) was performed to evaluate the lncRNA EPB41L4A-AS2 expression level in tissues and cell lines. Proliferating cell nuclear antigen (PCNA) protein level was determined by western blot assay. CCK8 assay, EdU assay, flow cytometry (FCM) and transwell assay were performed to access cell proliferation, apoptosis and invasion. EPB41L4A-AS2 expression was significantly downregulated in cancer tissues and cells compared with the adjacent tissues and normal cells (P<0.05). After cells were transfected with pcDNA3.1-EPB41L4A-AS2, cell viability and PCNA protein level was decreased, and cells were arrested in the G0/G1 phase with higher apoptosis rate. Transwell assay showed that over-expressed EPB41L4A-AS2 could reduce cells invasion ability. Expression of low levels of EPB41L4A-AS2 is associated with poor survival in NSCLC and the over-expression of lncRNA EPB41L4A-AS2 inhibits NSCLC cell proliferation, invasion and promote cell apoptosis.