Objective To investigate the relationship between the levels of hypoxia inducible factor-1α (HIF-1α), thymidine phosphorylase (TP), vascular endothelial growth factor (VEGF) and lymph node metastasis in breast tumor. To provide reference for clinical practice. Methods Eighty patients with breast cancer who received surgical treatment in Qingdao Chengyang People’s Hospital from September 2017 to September 2022 were selected as the research objects for retrospective analysis, and the levels of HIF-1α、 TP and VEGF in breast cancer tissue resected surgically and normal adjacent breast tissue were detected to explore the relationship between them and clinicopathological characteristics. Results The positive expression rates of HIF -1α, TP and VEGF in breast cancer tissues were higher than those in adjacent tissues (P<0.05). There was no significant difference in the positive expression rates of HIF -1α, TP and VEGF between patients in catheter infiltration group and non-invasive group (P>0.05).The positive expression rates of HIF -1α, TP and VEGF in the well differentiated group and the middle differentiated group were lower than those in the poorly differentiated group (P<0.05). There was no significant difference in the positive expression rates of HIF -1α, TP and VEGF between the well differentiated group and the middle differentiated group (P>0.05). The positive expression rates of HIF -1α, TP and VEGF in stage I and stage Ⅱ group patients were lower than those in stage Ⅲ group (P<0.05). There was no significant difference in the positive expression rates of HIF -1α, TP and VEGF between the patients in stage I and stage Ⅱ group (P>0.05). The positive expression rates of HIF -1α, TP and VEGF in patients with metastasis were higher than those in patients without metastasis (P<0.05). Conclusion HIF-1α、 TP and VEGF were positive in cancer tissues, and were closely related to lymph node metastasis.