AKR1B1 controls intracellular PGF2α levels and mediates the NF-κB pathway. (A) PGF2α biosynthesis pathway. (B and C) The level of PGF2α was measured in MDA-MB231 and SUM159 cells with stable empty vector (Ctr) or knockdown of AKR1B1 expression (B), as well as T47D and MCF7 cells with stable empty vector or AKR1B1 expression (C). The level of PGF2α is shown in the bar graph. (D and E) Migratory ability (D) and invasiveness (E) of MDA-MB231 and SUM159 cells treated with or without 100 µM PGF2α were analyzed. The percentage of migratory and invasive cells is shown in the bar graph. (F) Expression of AKR1B1, RelA, and IKBα was analyzed by Western blotting in MDA-MB231 with stable empty vector or knockdown of AKR1B1 expression (left), as well as T47D cells with stable empty vector or AKR1B1 expression (right). (G and H) NF-κB reporter activity was examined in MDA-MB231 and SUM159 cells with stable empty vector or knockdown of AKR1B1 expression (G), as well as T47D and MCF7 cells with stable empty vector or AKR1B1 expression (H). After 48 h, luciferase activities were normalized and determined. Data are mean ± SD in three separate experiments. *, P < 0.01 by Student’s t test.