7,3′,4′-Trihydroxyisoflavone modulates multidrug resistance transporters and induces apoptosis via production of reactive oxygen species

Highlights: ► We explore the mechanisms of 7,3′,4′-trihydroxyisoflavone to potentiate cytotoxicity of epirubicin. ► This isoflavone decreases the activity of hMDR1 promoter region in HeLa cells. ► This combination causes a ROS-dependent suppression of MDR transporters. ► This combination initiates a p53-mediated activation of the mitochondrial pathway of apoptosis. ► 7,3′,4′-Trihydroxyisoflavone has the potential to reduce anticancer drug dosage and corresponding side effects. -- Abstract: The development of multidrug resistance (MDR) to conventional chemoradiation therapy usually leads to failure in treating cervical cancer. This study aims to explore the effects and mechanisms of 7,3′,4′-trihydroxyisoflavone (7,3′,4′-THIF), one of the major metabolites of daidzein, on potentiating cytotoxicity of epirubicin (Epi), an anticancer drug in human cervical cancer HeLa cells. The cytotoxicity of Epi remarkably increased when it was combined with 7,3′,4′-THIF. The cotreatment increased the reactive oxygen species (ROS) levels, including hydrogen peroxide and superoxide free radicals. 7,3′,4′-THIF was shown to down-regulate the MDR1 promoter region composed of the elements of AP1, GC-box, and Y-box, as demonstrated by a luciferase assay. A negative regulation of hMDR1 gene with multiple transcription factors by this isoflavone may provide a novel molecular mechanism for MDR modulation. The mRNA expressions of MDR1, MDR-associated protein (MRP) 1, and MRP2 for the combined treatment were significantly lower than those of the Epi treatment. This result implies that MDR transporter-mediated Epi resistance is inhibited at various degrees by the addition of 7,3′,4′-THIF. This isoflavone significantly enhanced intracellular Epi accumulation in HeLa cells. 7,3′,4′-THIF and/or Epi triggered apoptosis through the upregulation of p53, Bax, and caspase-9. Apoptosis induction was also confirmed by the reduced mitochondrial membrane potential, increased sub-G1 and G2/M phases, nuclear DNA fragmentation, and chromatin condensation. Our findings demonstrate for the first time that 7,3′,4′-THIF causes cell death in human cervical cancer cells through the ROS-dependent suppression of MDR transporters and p53-mediated activation of the intrinsic mitochondrial pathway of apoptosis. Thus, 7,3′,4′-THIF has the potential to enhance the activity of a broad range of cancer chemotherapeutics in the MDR spectrum with the advantage of reducing adverse effects.