Objective: To investigate the relationship between the expression level of protein tyrosine kinase STYK1 and cisplatin resistance in non-small cell lung cancer (NSCLC). Methods: Kaplan-Meier Plotter/GEPIA database was used to analyze the correlation between the expression of STYK1 and the prognosis of NSCLC patients after chemotherapy. Eighty-one NSCLC patients who received docetaxel combined with cisplatin were randomly enrolled. The expression level of STYK1 was detected by immunohistochemical staining, and the relationship between STYK1 expression and the effect of chemotherapy was analyzed. The expression of STYK1 in lung cancer A549 and cisplatin resistant A549/DDP cells was detected by real-time fluorescent quantitative PCR (qRT-PCR) and Western blot. A549/DDP cells with low expression of STYK1 were constructed. Under the stimulation with different concentrations of cisplatin, the cell activity was detected by CCK-8 method and the growth curve was plotted. Results: Bioinformatics analysis showed that the overall survival and progress-free survival of patients with high STYK1 expression were significantly lower than those with low expression of STYK1, and the expression level of STYK1 and PI3K was correlated. Immunohistochemical staining showed that the high expression rate of STYK1 in the chemotherapy non-remission group was significantly higher than that in the chemotherapy remission group (71.4% vs 46.9%, P<0.05). qRT-PCR and Western blot results showed that the expression level of STYK1 in A549/DDP cells was higher than that in A549 cells, and the down-regulation of STYK1 expression could significantly inhibit the activity and proliferation of A549/DDP. Conclusion: The expression level of STYK1 is correlated with the effect of cisplatin chemotherapy in NSCLC, and it is expected to become a new target for predicting and improving the effectiveness of chemotherapy in lung cancer.