The Mechanism of Luteolin Promoting Cuprotosis and Enhancing Cisplatin Sensitivity of Breast Cancer Cells by Regulating PI3K/AKT/GSK-3β Signaling Pathway
BI Chao, SHI Xiaoguang, HU Junsheng, LAI Rui, ZHANG Juan, HUO Qiaoling, LIU Man*
The aim of this study was to investigate the effect of luteolin on the sensitivity of BC (breast cancer) cells to DDP (cisplatin) and its mechanism of promoting copper death by regulating PI3K/AKT/GSK-3β signaling pathway. The cisplatin-resistant breast cancer cell line MDA-MB-231/DDP-R was established by continu ous induction with low concentration. MTT assay was used to detect the effects of different concentrations of cisplatin and breast cancer on the inhibition rate of MDA-MB-231 and MDA-MB-231/DDP-R. MDA-MB-231/DDP-R cells were divided into five groups: control group, cisplatin group, luteolin group, cisplatin+luteolin group and 740Y-P group. The cell invasion, migration and apoptosis levels were detected by Transwell assay, wound healing assay and f low cytometry, respectively. The intracellular Cu2+ level was measured, and the expression levels of proteins related to cuproptosis and the PI3K/AKT/GSK-3β signaling pathway were detected by Western blot. Nude mice xenografts were constructed to detect tumor volume and weight. The results showed that the inhibitory rates of cisplatin on both MDA-MB-231 and MDA-MB-231/DDP-R cells increased significantly with rising cisplatin concentrations (P<0.05). Compared with MDA-MB-231 cells, the inhibitory effects of cisplatin and luteolin on MDA-MB-231/DDP-R cells were significantly lower (P<0.05). The cisplatin+luteolin group exhibited a significantly higher cell inhibitory rate compared to the cisplatin group alone (P<0.05). Compared with the Control group, the cisplatin, luteolin, and cisplatin+luteolin groups showed significant reductions (P<0.05) in the number of invasive cells, cell migration rate, ATP7A protein expression, the ratios of p-PI3K/PI3K, p-AKT/AKT, and p-GSK-3β Ser9/GSK-3β, as well as in nude mouse tumor volume and weight. Conversely, the cell apoptosis rate, intracellular Cu⁺ concentration, and the expression levels of DLAT and FDX1 proteins were significantly increased (P<0.05). Compared to either the cisplatin group or the luteolin group alone, the cisplatin+luteolin group demonstrated significantly lower values (P<0.05) for the number of invasive cells, cell migration rate, ATP7A protein expression, the p-PI3K/PI3K, p-AKT/AKT, and p-GSK-3β Ser9/GSK-3β ratios, and tumor volume and weight. In contrast, the cell apoptosis rate, Cu⁺ concentration, and DLAT and FDX1 pro tein expression levels were significantly higher (P<0.05). Compared with the cisplatin+luteolin group, the 740Y-P group exhibited significant increases (P<0.05) in the number of invasive cells, cell migration rate, ATP7A protein expression, the p-PI3K/PI3K, p-AKT/AKT, and p-GSK-3β Ser9/GSK-3β ratios, and tumor volume and weight. Meanwhile, the cell apoptosis rate, Cu⁺ concentration, and DLAT and FDX1 protein expression levels were signifi cantly decreased (P<0.05). luteolin can induce cuprotosis by regulating the PI3K/AKT/GSK-3β signaling pathway, thereby enhancing the sensitivity of breast cancer cells to cisplatin.
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