目的:探讨从黄花香茶菜中提取的新型二萜化合物黄花香茶菜甲素(Sculponeatin A,stA)诱导乳腺癌铁死亡的作用机制.方法:使用实时无标记细胞分析技术、克隆形成实验和共聚焦高内涵成像分析评估stA对乳腺癌细胞增殖的影响;利用活性氧检测试剂盒和流式细胞术,以及谷胱甘肽检测试剂盒、丙二醛检测试剂盒、亚铁离子检测试剂盒等探究stA对乳腺癌细胞铁死亡的影响;利用Western blot检测stA对铁死亡上游调控因子ETS1(E26 transformation specific 1)蛋白表达的作用;通过分子对接,微量热泳动实验以及基于靶点稳定性的药物亲和反应检测stA与ETS1 的结合作用.结果:stA在纳摩尔级剂量下抑制乳腺癌细胞的生长,并改变细胞的形态;stA能够诱导乳腺癌细胞铁死亡;stA通过下调ETS1 的蛋白表达来诱导乳腺癌细胞发生铁死亡;stA与ETS1 有直接结合能力.结论:stA能够直接结合ETS1,通过下调ETS1 表达而诱导乳腺癌xCT途径铁死亡.
Mechanism of Sculponeatin A Inducing Ferroptosis in Breast Cancer Cells by Targeting ETS1
Objective To investigate the mechanism of sculponeatin A(stA),a novel diterpenoid compound extracted from Isodon sculponeatus,in inducing ferroptosis in breast cancer cells.Methods The effect of stA on the proliferation of breast cancer cells was evaluated by real-time cell analysis,colony formation assay,and high-content imaging.Its impact on the ferroptosis of breast cancer cells was determined by flow cytometry,glutathione,malondialdehyde,and iron determination assay.Western blot was conducted to assess the influence of stA on the protein expression of E26 transformation specific 1(ETS1),an upstream regulator of ferroptosis.The binding of stA and ETS1 was examined through molecular docking,mi-croscale thermophoresis,and drug affinity-responsive target stability.Results stA inhibited the proliferation of breast canc-er cells and induced changes in cell morphology at a molar concentration.Furthermore,stA was found to induce ferroptosis in breast cancer cells by down-regulating ETS1.It was also determined that stA had a direct affinity with ETS1.Conclu-sion These findings indicate that stA directly binds to ETS1 and induces ferroptosis in breast cancer cells through the down-regulation of ETS1 in the xCT pathway.
Breast cancerSculponeatin AE26 transformation specific 1(ETS1)xCTFerroptosis
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