Santolina chameacyparissus (Santo) is an evergreen plant which is traditionally used for its anti-inflammatory effects in various diseases. In this study, we aimed to explore the effects of Santo in non-small cell lung cancer cells. We extracted volatile oil from the plant and evaluated cytotoxicity, apoptosis, and motility effects of the extract on two non-small cell lung cancer (NSCLC) cell lines; one is a patient derived and the other one is a commercially available A549 cells. We also identified its components via GC/MS and investigated possible targets of the major components of the plant using qPCR and docking studies. Cytotoxicity tests showed dose dependent cell killing activity and flow cytometry assays exposed apoptotic effects of Santo. The essential oil also remarkably decreased migration rate of A549 cells, therefore we evaluated the expression levels of epithelial to mesenchymal transition related genes E-cadherin and Vimentin ratio, ZEB1 and SNAIL and another motility related gene Ezrin. Santo did not change the expression of EMT related genes but decreased Ezrin levels. According to the results of the GS/MS analysis, Artemisia ketone and Camphor were identified as major molecules of the extract. Docking analysis also revealed that artemisia ketone, the major component of the Santo extract, potentially showed strong binding to the active site of ezrin protein and both artemisia ketone and camphor had ability to bind DNA. The results of the present study indicate that Santo and its components artemisia ketone and camphor are promising anti-cancer agents, and their potential in targeting DNA and oncogenic proteins in the lung cancer cells seems worth to focus on this plant in cancer related drug discovery science.