Figure 3.

Canagliflozin inhibits mitochondrial complex-I to limit cancer cell proliferation. (A) Extracellular acidification rate (ECAR, mOD/min) of PC3 cells 3 h post treatment with Canagliflozin (30 μM), Dapagliflozin (30 μM), or vehicle (n = 4, in quadruplicate). (B) Rate of oxygen consumption (pmol/s/10⁶ cells) through complex-I of permeabilized PC3 cells treated with increasing concentrations of Canagliflozin and Dapagliflozin (n = 4, in duplicate). (C) Rate of oxygen consumption (pmol/s/10⁶ cells) through complex-II of permeabilized PC3 cells treated with increasing concentrations of Canagliflozin and Dapagliflozin (n = 4, in duplicate). (D) Western blot analysis of NDI1 expression in PC3-pMXS and PC3-NDI1 cells. (E) Rate of lipogenesis (nmol of [H³-acetate] incorporation into fatty acid/mg of protein/hour) in PC3-pMXS and PC3-NDI1 cells treated with Canagliflozin (30 μM) for 4 h relative to the vehicle control (n = 3, in triplicate). (F) Cellular proliferation of PC3-PMXS and PC3-NDI1 cells treated with Canagliflozin or Phenformin and expressed relative to the vehicle controls for 48 h (n = 4, in quadruplicate). (G) Graphical representation of the proposed mechanism of Canagliflozin. Results are expressed as the mean and standard error of the mean (SEM). Vehicle versus treatment ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by one-way ANOVA for A, by two-way ANOVA for B. Canagliflozin versus Dapagliflozin #### = p < 0.0001 by two-way ANOVA for B. pMXS versus NDI1 * = p < 0.05, ** = p < 0.01 by t-test for E, by two-way ANOVA for F.