Our findings reveal that PKMYT1AR/miR-485-5p /PKMYT1 axis is important for cancer stem cell maintenance and NSCLC progression both in vitro and in vivo, suggesting that PKMYT1AR, miR-485-5p or PKMYT1 could be used as novel therapeutic targets in the future. The gene discussed is PKMYT1; the disease is non-small cell lung carcinoma.