Similarly, high SMAD4 methylation degree was significantly associated (P < 0.05) with high grade (74 ± 14.4 vs. 63.3 ± 12.01), tumor late stage (84.2 ± 17.4 vs. 64.5 ± 12.7), positive lymph node metastasis (75.1 ± 14.1 vs. 65.6 ± 13.4), positive ER (72 ± 15.5 vs. 68.2 ± 13.2), positive PR (72.8 ± 14.5 vs. 65.2 ± 13.3), and positive HER2 (72.3 ± 14.6 vs. 66 ± 13.4) as shown in Fig. 4a–f. Here, ESR1 is linked to metastatic malignant neoplasm in the lymph nodes.