Objective To detect the expression levels of Cbl-b, c-Cbl, and Cbl-c proteins of E3 ubiquitin ligase casitas b-lineage lymphoma (Cbl) family in non-small cell lung cancer (NSCLC), and to analyze their relationship with clinicopathological features and prognosis. Methods This was a prospective study. A total of 102 patients with NSCLC admitted to Hanzhong People's Hospital from January 2020 to January 2023 were selected, including 78 males and 24 females. They were (67.49±8.33) years old. The cancer-adjacent tissues removed by surgery were used as controls. The immunohistochemical staining was used to detect the expressions of Cbl-b, c-Cbl, and Cbl-c proteins in the cancer tissues and cancer-adjacent tissues. The expressions of Cbl-b, c-Cbl, and Cbl-c proteins in the cancer tissues of the patients with different clinicopathological characteristics (gender, age, smoking history, lesion site, TNM stage, differentiation grade, pathological type, and pleural invasion) were compared. The Cox regression model was used to analyze the influencing factors of the patients' prognosis. Kaplan-Meier curves were drawn to analyze the relationship between the expressions of Cbl-b and Cbl-c proteins in the cancer tissues and NSCLC prognosis. t and χ2 tests were applied. Results The positive expression rates of Cbl-b, c-Cbl, and Cbl-c proteins in the cancer tissues were higher than those in the cancer-adjacent tissues [52.94% (54/102) vs. 37.25% (38/102), 63.73% (65/102) vs. 43.14% (44/102), and 67.65% (69/102) vs. 39.22% (40/102)], with statistical differences (χ2=5.068, 8.688, and 16.568; all P<0.05). The positive expression rates of Cbl-b, c-Cbl, and Cbl-c proteins in the cancer tissues of the patients with smoking history, TNM stage Ⅲ/Ⅳ, undifferentiated/low differentiation, and pleural invasion were higher than those of the patients without smoking history and with TNM stage I/Ⅱ, moderate/high differentiation, and no pleural invasion [smoking history: 62.32% (43/69) vs. 33.33% (11/33), 73.91% (51/69) vs. 42.42% (14/33),and 78.26% (54/69) vs. 45.45% (15/33); TNM stage: 76.00% (19/25) vs. 45.45% (35/77), 88.00% (22/25) vs. 55.84% (43/77), and 92.00% (23/25) vs. 59.74% (46/77); differentiation: 65.22% (30/46) vs. 42.86% (24/56), 78.26% (36/46) vs. 51.79% (29/56), and 82.61% (38/46) vs. 55.36% (31/56); pleural invasion: 80.00% (20/25) vs. 44.16% (34/77), 88.00% (22/25) vs. 55.84% (43/77), and 92.00% (23/25) vs. 59.74% (46/77)], with statistical differences (χ2=7.528, 9.575, 10.978, 7.068, 8.442, 8.974, 5.068, 7.658, 8.570, 9.733, 8.442, and 8.974; all P<0.05). The Cox regression analysis showed that smoking history, TNM stage Ⅲ, undifferentiated/low differentiation, pleural invasion, and Cbl-b protein expression and Cbl-c protein expression in the cancer tissues were the influencing factors of the patients' prognosis (all P<0.05). The progression-free survival time of the Cbl-b positive group was 27 months (95%CI 22.602-31.398 months), and that of Cbl-b negative group was 40 months (95%CI 28.810-51.190 months); the overall comparison of Kaplan-Meier survival curves showed a statistical difference (χ2=6.709, P=0.010). The progression-free survival time of the Cbl-c positive group was 28 months (95%CI 23.786-32.214 months), and that of the Cbl-c negative group was 39 months (95%CI 29.892-48.108 months); the overall comparison of Kaplan-Meier survival curves showed a statistical difference (χ2=4.605, P=0.032). Conclusions Cbl-b, c-Cbl, and Cbl-c proteins are highly expressed in NSCLC. All the three are related to smoking history, TNM stage, differentiation degree, and pleural invasion. Among them, Cbl-b and Cbl-c are influencing factors of prognosis of patients with NSCLC.