乙氧基血根碱通过激活AMPK信号通路诱导自噬抑制三阴性乳腺癌增殖的作用机制研究

Mechanism of Ethoxysanguinarine Inhibiting Triple-negative Breast Cancer Proliferation by Activating AMPK Signaling Pathway and Inducing Autophagy

目的:探讨乙氧基血根碱(ethoxysanguinarine,Eth)能否通过AMPK信号通路诱导自噬抑制三阴性乳腺癌(triple negative breast cancer,TNBC)的增殖。方法:通过MTT法、台盼蓝拒染法及集落形成实验检测Eth对2种不同TNBC细胞系(MDA-MB-231和MDA-MB-436)体外增殖能力的影响;通过Western blot和免疫荧光实验检测Eth对2种不同的TNBC细胞系自噬的影响;通过Western blot检测Eth对AMPK/mTORC1及其下游信号通路的影响;通过分子对接和免疫共沉淀分析Eth与AMPK的相互作用关系。结果:Eth在低微摩尔浓度剂量下能够显著抑制TNBC细胞的增殖;Eth能够诱导TNBC细胞发生自噬;Eth能够在TNBC细胞中激活AMPK/mTORC1信号通路;Eth与AMPK存在直接的结合作用。结论:Eth通过直接靶向活化AMPK诱导TNBC细胞发生自噬进而抑制TNBC细胞增殖。Eth有望作为一种潜在的抗肿瘤候选药物用于TNBC患者的治疗。

Objective This article aims to investigate whether ethoxysanguinarine(Eth)can induce autophagy through AMPK signaling pathway to inhibit the proliferation of triple negative breast cancer(TNBC).Methods The effect of Eth on the proliferation ability of two different TNBC cell lines(MDA-MB-231 and MDA-MB-436)in vitro was detected by MTT method,trypan blue exclusion method and colony formation assay.The effect of Eth on autophagy of two different TNBC cell lines was detected by Western blot and immunofluorescence assay.The effect of Eth on AMPK/mTORC1 and its downstream signal pathway was detected by Western blot.The interaction between Eth and AMPK was analyzed by molecular docking and immunocoprecipitation.Results Eth could significantly inhibit the proliferation of TNBC cells at low micromolar concentrations.Eth could induce autophagy in TNBC cells.Eth could activate AMPK/mTORC1 signaling pathway in TNBC cells.Eth had a direct binding effect with AMPK.Conclusion Eth inhibits the proliferation of TNBC cells by directly targeting and activating AMPK.Therefore,Eth is expected to be a potential antitumor drug candidate for the treatment of TNBC patients.