Abstract: OBJECTIVE To investigate the correlation between high-risk human papillomavirus (HPV) infection, P53 protein and breast cancer. METHODS From Jan. 2014 to Jan. 2016, a total of 60 cases of breast cancer patients were prospectively collected as study group, while 60 patients with cyclomastopathy were enrolled in this study as control group. The high-risk gene types of HPV infection and expression of P53 protein in both breast cancer tissue and cyclomastopathy tissue were detected. Moreover, the association between high-risk HPV infection and P53 protein and the clinical parameters of breast cancer were analyzed. RESULTS The infection rates of HPV16 and HPV18 in study group were 20.00% and 23.33%, which were significantly higher than those of control group of 3.33% and 3.33% (P<0.05). There was no significant difference in the infection rates of 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68 HPV genotypes between the two groups. The positive rate of P53 protein in HPV positive breast cancer patients was significantly lower. The lymph node metastasis rate of HPV positive patients was significantly higher than that of HPV negative patients. There was no significant correlation between HPV infection and TNM stage and tumor size. CONCLUSION High-risk HPV infection is associated with P53 protein and lymph node metastasis in patients with breast cancer.