Objective To analyze the relationship between integrin α3 (ITGA3) and serum interferon gamma (IFN-γ) and clinicopathological features and prognosis of patients with non-small cell lung cancer. Methods 86 patients with non-small cell lung cancer from October 2020 to October 2022 were selected as the experimental group, and 36 patients with benign lesions were selected as the control group. The serum ITGA3 and IFN-γ levels in patients with different groups and different prognosis were compare. The relationship between positive and negative levels and clinicopathological characteristics of patients was analyzed. Cox proportional hazard regression model was used to analyze the effects of ITGA3 and IFN-γ levels on survival and prognosis. Results The positive expression rate of IFN-γ in the experimental group was 61.63%, which was significantly higher than 13.89% in the control group (χ2=23.190, P<0.05). The positive expression rate of ITGα3 in the experimental group was 55.81%, significantly higher than that in the control group (11.11%) (χ2=20.736, P<0.05). The positive expression of IFN-γ was not correlated with gender, age and differentiation degree (P>0.05), but was significantly correlated with smoking history, TNM stage, pathological type and lymph node metastasis (P<0.05). The positive expression of ITGα3 was not correlated with sex, age, TNM stage and differentiation degree (P> 0.05), but was significantly correlated with smoking history, pathological classification and lymph node metastasis (P<0.05). The expressions of IFN-γ and ITGα3 in death group were significantly higher than those in non-death group (P<0.05). Cox regression model analysis showed that positive expressions of IFN-γ and ITGα3 were independent risk factors for survival and prognosis of NSCLC patients (P<0.05). Conclusion ITGA3 and serum IFN-γ are significantly correlated with smoking history, pathological classification and lymph node metastasis. Positive expressions of IFN-γ and ITGα3 are independent risk factors affecting the survival and prognosis of patients with NSCLC. Observation of their levels is beneficial to the diagnosis of the disease and is helpful for clinical evaluation of treatment effect