Fig 2: Immunohistochemical analysis of thymidylate synthase (TYMS) protein expression in (A) breast cancer and adjacent normal breast tissues, (B) triple‐negative breast cancer (TNBC) and non‐TNBC breast cancer tissues. Association of TYMS protein expression with (C) ER status, (D) PR status, (E) KI67 status and (F) HER2 status. (G) Association of TYMS protein expression with OS of breast cancer patients

Fig 3: A, Expression of thymidylate synthase (TYMS) mRNA in breast cancer and adjacent normal breast tissues (P < .0001). B, Expression of TYMS mRNA in triple‐negative breast cancer (TNBC) and non‐TNBC tissues (P < .0001)

Fig 4: Association of thymidylate synthase (TYMS) mRNA expression with (A) overall survival (OS) of breast cancer patients, (B) disease‐free survival (DFS) of breast cancer patients and (C) recurrence‐free survival (RFS) of breast cancer patients

Fig 5: Trapping efficiency of FdUMP by TS is higher in HCT116RF10 cells than in parental HCT116 cells. (A) Dynamics of the TS protein in TYMS-silenced HCT116RF10 cells and HCT116 cells after treatment with 5-FU. At 48 h after transfection with TYMS-targeted siRNA or nonsilencing siRNA, the cells were treated with 5-FU 100 μM for the indicated treatment time, and whole-cell lysates were prepared. The protein expression of TS and β-actin was measured by Western blotting analysis. The data are representative of at least three independent experiments. NSsi, nonsilencing siRNA; TSsi, TYMS-targeted siRNA; (B) TS protein level in HCT116RF10 cells and HCT116 cells. The levels of total TS, i.e., the active free-TS form and the inactive FdUMP-TS form, are indicated by the ratio of TS density to β-actin density for each treatment relative to the value for the NSsi-transfected parental HCT116 cells without 5-FU. The results represent the average of three independent experiments and the error bars show the ±SE of triplicate experiments. White bar, free-TS form; gray bar, FdUMP-TS form. Student’s t-test, * p < 0.05 and ** p < 0.01, one-way ANOVA, p < 0.001 (for total TS levels of all groups), and p < 0.05 (for FdUMP-TS levels of all groups).