〔Abstract〕 To explore the effect of transcription factor ZNF174 on the stemness of non-small cell lung cancer (NSCLC) . Methods StemChecker online analysis was employed to assess the impact of ZNF174 on tumor stemness markers . The ZNF174 knockdown plasmid was constructed via restriction enzyme digestion and enzyme- linked assembly , while the ZNF174 overexpression plasmid was generated through homologous recombination . Lentiviral infection established stable cell lines with ZNF174 knockdown or overexpression . Tumor stem cell sphe- roid formation assays were conducted to evaluate stemness maintenance capacity. RNA-seq and quantitative real- time polymerase chain reaction (qPCR) were utilized to detect effects on key stemness gene expression . Results The transcription factor ZNF174 promoted cellular stemness in non-small cell lung cancer (NSCLC) . It promoted the expression of key stemness genes : B-cell-specific moloney murine leukemia virus insertion site 1 (BMI1) , ac- etaldehyde dehydrogenase 2 (ALDH2) , SRY-box transcription factor 2 (SOX2) , recombinant high mobility group AT hook protein 2 (HMGA2) , and snail family transcriptional repressor 1 (Snail1) and enhanced tumor spheroid formation in NSCLC stem cells . Conclusion ZNF174 promotes the expression of tumor stemness genes in NSCLC and may become a novel therapeutic and prognostic target for NSCLC .