Sulfasalazine induces ferroptosis of breast cancer ZR-75-1 cells and its mechanism / 肿瘤

ABSTRACT

Objective: To observe the activation effect of sulfasalazine on ferroptosis of breast cancer ZR-75-1 cells, and to explore its possible mechanism. Methods: The different concentrations of sulfasalazine (0, 1.0, 0.5, 1.0, and 2.0 mmol/L) were used to treat ZR-75-1 cells. The morphological change of ZR-75-1 cells was observed under an inverted microscope. The proliferation inhibitory rate of ZR-75-1 cells was detected by CCK-8 method. The expression levels of glutathione peroxidase 4 (GPX4) and cysteine-glutamate antiporter subunit (xCT) proteins in ZR-75-1 cells were detected by Western blotting. The fluorescent probe was used to label the ZR-75-1 cells treated with different concentrations of sulfasalazine, then the change of reactive oxygen specie (ROS) level (as indicated by fluorescence intensity) was detected by FCM method. The expression level of divalent metal transporter 1 (DMT1) mRNA was detected by real-time fluorescent quantitative PCR. Results: Sulfasalazine caused the morphological changes and death of ZR-75-1 cells. The low doses of sulfasalazine had little effect on the proliferation of ZR-75-1 cells (P > 0.05), but the high concentrations of sulfasalazine inhibited cell proliferation significantly (P < 0.05). The high concentrations of sulfasalazine inhibited the expressions of GPX4 and xCT proteins in ZR-75-1 cells (both P < 0.05), and also promoted the accumulation of intracellular ROS (P < 0.05). Sulfasalazine increased the expression of DMT1 mRNA in ZR-75-1 cells (P < 0.05). Conclusion: Sulfasalazine can increase the accumulation of ROS by inhibiting the expressions of GPX4 and xCT proteins to induce the ferroptosis of ZR-75-1 cells. In which, the activation of DMT1 expression may be one of the mechanism.