Figure 1 Immunohistochemical staining for CD44, CD24, and DAPI (×400). Table 2 Proportion of all patients and patients with recurrence/metastasis and CD44/CD24 data with CD44+/CD24-/low tumor cells   n All cases (%) P n Recurrence/metastatic cases (%) P* Age (years) < 50 74 18.34 ± 2.70 0.444 34 24.91 ± 3.79 0.022 ≥ 50 73 15.45 ± 2.66   38 13.20 ± 3.32

  Tumor size T1 47 15.78 ± 2.86 0.224 15 13.19 ± 3.53   T2 76 20.12 ± 2.90   44 23.78 ± 3.68   T3 + T4 17 10.27 ± 4.46   13 11.83 ± 6.60 0.152 Lymph node involvement Absent 32 8.66 ± 2.70 0.026 18 10.00 ± 3.77 0.075 Present 115 19.20 ± 2.29   54 21.53 ± 3.19   TNM stage I + II 70 15.87 ± 2.63 0.500 33 16.88 ± 3.74 0.368 this website III + IV 77 18.49 ± 2.81   39 21.73 ± 3.79   ER expression Negative 90 16.49 ± 2.47 0.845 47 18.92 ± 3.17 0.944 Positive 57 17.26 ± 3.07   25 19.32 ± 4.81   PR expression Negative 83 13.09 ± 2.41 0.038 43 14.63 ± 3.06 0.046 Positive 64 21.06 ± 2.98   29 25.32 ± 4.51

  Her2 expression Negative 77 16.18 ± 3.03 0.566 38 17.36 ± 4.17 0.441 Positive 70 18.47 ± 2.61   34 21.57 ± 3.47   Basal-like feature † Absent 108 18.44 ± 2.24 0.143 49 11.70 ± 4.07 0.050 Present 39 11.93 ± 3.66   23 22.66 ± 3.30   Recurrence or metastasis Absent 75 14.26 ± 2.72 0.246       Present 72 18.73 ± 2.58         Lesions in recurrence/metastatic patients Primary       56 15.39 ± 2.63 0.014 Secondary AZD9291 ic50       16 30.41 ± 6.46   * Calculated by t tests. ER, estrogen receptor; PR, progesterone receptor; Her2, human epidermal growth factor receptor 2. † Immunohistochemically negative for both SR and Her2. Association of CD44+/CD24- phenotype with steroid receptor status Of the 121 samples with CD44/CD24 data, 56 (46.2%) were positive for PR expression. CD44+/CD24- status was significantly correlated with strong PR staining in all patients (P = 0.038) and in samples from patients with recurrence or metastasis (P = 0.046). Interestingly, although ER expression was observed in 50 of the 121 (41.3%) patients with CD44/CD24 data,

the presence of CD44+/CD24- tumor cells was not significantly correlated with positive ER expression in all patients and in GNA12 patients with recurrence or metastasis. Association of CD44+/CD24- phenotype with basal-like feature We found that the proportion of CD44+/CD24- tumor cells was similar in breast cancer samples with and without basal-like features (11.93% versus 18.44%, p = 0.143). However, in samples from patients with tumor recurrence or metastasis, the proportion of CD44+/CD24- tumor cells was significantly higher in breast cancer tissue with basal-like features than in tissue without such features (22.66% versus 17.70%, p = 0.05). Association of CD44+/CD24- phenotype with DFS and OS: univariate analysis and multivariate analysis The results of univariate analyses of the associations between each individual predictor and DFS are shown in Table 3. The proportion of CD44+/CD24-/low tumor cells (P = 0.002), PR status (P = 0.