The aim of the present study was to investigate the effect of CpG-oligodeoxynucelotides (CpG-ODN) on the proliferation of the A549 human lung adenocarcinoma cell line and the expression of Runt-related transcription factor 3 (Runx3) and investigate the association between the toll-like receptor 9 (TLR9) signaling pathway and Runx3 expression during A549 cell proliferation. Here, TLR9 is linked to lung adenocarcinoma.