中国生物工程杂志

Objective: To explore the effects of miR-324-3p on prostate cancer (PCa) cell ferroptosis and its underlying mechanism. Methods: qRT-PCR was performed to detect the expression of miR-324-3p and glutathione peroxidase 4 (GPX4) mRNA in PCa tissues, matched adjacent tissues, and cell lines. Western blot was employed to examine the protein level of GPX4 in PCa cell lines and normal prostate epithelial cells. CCK-8 assay was used to evaluate cell proliferation. The levels of glutathione (GSH), lipid oxidation, and reactive oxygen species (ROS) were determined using GSH detection assay kit, lipid peroxidation MDA assay kit and DCFH-DA fluorescent probe assay. Dual-luciferase reporter gene was applied to verify the interaction of miR-324-3p and GPX4. Results: The expression of miR-324-3p in PCa tissues was lower than that in the corresponding paracancerous tissues, and the expression of miR-324-3p was downregulated in PCa cell lines compared with normal prostate epithelial cells. GPX4 was highly-expressed in PCa tissues and cell lines in comparison with the corresponding paracancerous tissues and normal prostate epithelial cells. Dual-luciferase reporter results showed that miR-324-3p was directly targeted to negatively regulate GPX4. Overexpression of miR-324-3p significantly inhibited PCa cell proliferation, reduced GSH production and enhanced the levels of lipid oxidation and ROS; while treatment with the ferroptosis inhibitor Fer-1 or GPX4 reversed the promotion effect of miR-324-3p on ferroptosis in PCa cells. Conclusion: miR-324-3p promoted ferroptosis in PCa cells by targeting negative regulation of GPX4 expression and thus exerted anticancer effects on PCa.