These were verified by the log-rank test. A, B. Patients with detectable plasma ESR1 mutations (P < 0.0001) (A) and PIK3CA mutations (P = 0.0034) (B) showed significantly shorter duration of ET effectiveness in 69 MBC patients. C., D. Tracking analysis of cfDNA ESR1 mutations and PIK3CA mutations in 52 breast cancer patients with longitudinal samples. Patients were grouped into those who did not have any ESR1 or PIK3CA mutations over the course of treatment, those in whom ESR1 or PIK3CA mutations were maintained or acquired, and those in whom ESR1 or PIK3CA mutations disappeared in cfDNA. C. Patients in the loss of cfDNA ESR1 mutations group had a longer duration of ET effectiveness than patients with acquired or maintained numbers of cfDNA ESR1 mutations, but had a shorter duration of ET effectiveness than patients without mutations over the course of treatment (P < 0.0001). D. There was no statistically significant differences in these three groups; no PIK3CA mutations during treatment group, the loss of cfDNA PIK3CA mutations group, and the acquired or maintained numbers of cfDNA PIK3CA mutations group. Abbreviations: ET, endocrine therapy; cfDNA, cell-free DNA; MBC, metastatic breast cancer.