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| MAC30 Regulating Ferroptosis Mediated by JNK/Nrf2/HO-1 Pathway to Promote Apoptosis in Breast Cancer Cells |
| LUO Yongsheng, SUO Yueer, LIANG Weiping, et al |
| Jilin Vanguard Hospital, Jilin Changchun 130000, China |
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Abstract Objective: To investigate the effect of MAC30 on breast cancer cell apoptosis through the regulation of ferroptosis mediated by JNK/Nrf2/HO-1 pathway. Methods: The mRNA and protein expression levels of MAC30 were detected in normal breast cells and various breast cancer cell lines. MCF-7 cells were divided into four groups: the control group, the NC-shRNA group (MCF-7 cells transfected with MAC30-NC-shRNA), the MAC30-shRNA group (MCF-7 cells transfected with MAC30-shRNA), and the MAC30-shRNA+SP600125 group (MCF-7 cells transfected with MAC30-shRNA and treated with JNK inhibitor). RT-qPCR and Western blot were used to detect the mRNA and protein expression levels of MAC30 in each group of MCF-7 cells to verify the transfection efficiency. CCK-8 assay, clone formation experiment, and flow cytometry were used to detect cell viability, clone number, and apoptosis in each group. Kits were utilized to measure the levels of ROS, MDA, GSH, and Fe2+ within cells of each group. RT-qPCR and Western blot were conducted to detect the mRNA and protein expression levels of iron death-related genes (GPX4, SLC7A11) in cells of each group. Western blot was also used to detect the expression levels of apoptosis-related proteins (Caspase-3, Bax, Bcl-2) and proteins related to the JNK/Nrf2/HO-1 signaling pathway (JNK, p-JNK, Nrf2, HO-1) in cells of each group. Results: Compared to the control group and the NC-shRNA group, the MAC30-shRNA group exhibited decreased mRNA and protein expression levels of MAC30, cell viability, and clone number in MCF-7 cells (P<0.05). Additionally, the apoptosis rate of MCF-7 cells, expression levels of Caspase-3 protein and Bax/Bcl-2, mRNA and protein expression levels of ferroptosis-related genes, as well as the expression levels of p-JNK/JNK and Nrf2 (cytoplasmic) were increased (P<0.05). Conversely, the expression levels of Nrf2 (nuclear) and HO-1 were decreased (P<0.05). Compared to the MAC30-shRNA group, the MAC30-shRNA+SP600125 group exhibited increased levels of MAC30 mRNA and protein expression, cell viability, and clone number within MCF-7 cells (P<0.05). Additionally, there were decreases in the apoptosis rate, Caspase-3 protein levels, Bax/Bcl-2 expression ratios, mRNA and protein expression levels of ferroptosis-related genes, and expression levels of p-JNK/JNK and Nrf2 (cytoplasmic) (P<0.05). Conversely, the expression levels of Nrf2 (nuclear) and HO-1 increased (P<0.05). Conclusion: Knockdown of MAC30 may promote apoptosis in breast cancer cells by regulating iron death mediated through the JNK/Nrf2/HO-1 signaling pathway.
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2025, Vol. 31 
