Objective To discuss and analyze the effects and mechanisms of the Notch1/Hes1 pathway discussing and analyzing the influence and differentiation of lung cancer stem cell proliferation and differentiation.Methods Sixty lung cancer patients admitted to the Respiratory and Critical Care Medicine Department of Xinyu People's Hospital in Jiangxi Province from January 2020 to December 2022 were selected for this clinical practice study.Lung cancer stem cells A549(A549-SCs)were isolated and cultured from the patients,and flow cytometry was used to identify A549 CSCs for detection and screening of lung cancer stem cells.The selected lung cancer stem cells were divided into control group and observation group using random number table method,with 30 cases in each group.The control group was not treated with any treatment,while the observation group was treated with Temozolomide.The inhibition of lung cancer stem cell viability,impact on lung cancer stem cell migration ability,and mRNA levels of cell differentiation ability related factors Sox2 and Oct4 in A549 CSCs were compared between the two groups.Results The mRNA and protein expression levels,Notch1 and Hes1 protein expression levels of lung cancer stem cells in the observation group were lower than those in the control group,and the mRNA levels of cell differentiation ability related factors Sox2 and Oct4 in A549-CSCs in the observation group were lower than those in the control group,and the differences were statistically significant (P ＜0.05).Conclusion From the perspective of the Notch1/Hes1 pathway,Temozolomide,has an effect of inhibiting the proliferation and differentiation of lung cancer stem cells for lung cancer,and the above effect is achieved by inhibition of lung cancer stem cell Sox2,Oct4,and mRNA protein expression.