组蛋白去乙酰化酶泛抑制剂LBH589协同汉防己甲素通过ROS/MEK/ERK信号通路抗三阴性乳腺癌作用及机制研究

Triple-negative breast cancer (TNBC) lacks targeted drugs and generally resistant to the existing chemotherapeutics, which also poor in clinical prognosis with high postoperative recurrence rate. New and efficient treatment strategy be urgently needed. The advantages of drug combination in the field of anti-tumor drug development are worthy of our attention and research. Based on that histone deacetylase promotes the oncogenesis and development of TNBC, and p-glycoprotein is involved in the drug resistance of TNBC, our preliminary study found that, different dose of HDACs inhibitor LBH589 combined with tetrandrine-natural alkaloid extracted from Chinese herbal, show a synergistic effect on TNBC, including induce cell death, inhibit cancer stemness and cell metastasis. This combination treatment also activate ROS generation and downregulate level of p-MEK/p-ERK. However, the underlying mechanism that if the ROS activation and p-MEK/p-ERK downregulation, also P-gp and HDACs inhibition involved in the synergistic anti-tumor activity are unclear. TNBC cell lines and patients derived primary cells, as well as animal models and bioinformatics analysis will be applied to intensively explore the biological functions and the molecular signaling mechanisms affected by LBH589 and tetrandrine combination treatment. The intent of this study is to provide a novel strategy of drug combination therapy with high-efficiency and low toxicity for TNBC.

三阴性乳腺癌（TNBC）缺乏临床靶向治疗方法，且对现有化疗药物有普遍耐药性，患者预后差、术后复发率高，目前迫切需要开发新的有效治疗策略。多药联合在抗肿瘤药物开发领域的优势值得关注。基于组蛋白去乙酰化酶促进TNBC发生发展，且P-糖蛋白参与TNBC耐药机制，我们初步研究发现，不同浓度组合的HDAC泛抑制剂LBH589联合天然中草药提取生物碱汉防己甲素有协同诱导TNBC细胞死亡，抑制肿瘤干细胞和EMT潜力，且能显著激活ROS水平并下调p-MEK/p-ERK水平。但该信号通路和LBH589的HDAC抑制作用及汉防己甲素的P-gp抑制作用是否参与协同抗TNBC等具体作用机理还不清楚。本研究将在TNBC细胞系和临床肿瘤原代细胞及动物模型水平，结合生物信息学分析，深入探究LBH589协同汉防己甲素抗TNBC的作用方式和分子信号调控机制，为TNBC开发高效低毒的药物联合治疗新策略提供理论依据。

三阴性乳腺癌（TNBC）恶性程度高，缺乏临床靶向治疗方法，且对现有化疗药物有普遍耐药性，患者预后差、术后复发率高，目前迫切需要开发新的有效治疗策略。高效低毒的多药联合治疗方案在抗肿瘤药物开发领域的优势值得关注。我们初步研究发现，不同浓度组合的HDAC泛抑制剂LBH589联合天然中草药提取生物碱汉防己甲素有协同抗TNBC作用，但具体作用机理不清楚。本研究在TNBC细胞体外体系结合生物信息学分析，探究了LBH589协同汉防己甲素抗TNBC的作用方式和分子信号调控机制。研究结果发现，不同浓度组合的LBH589 联合汉防己甲素可以通过不同的作用功能发挥协同对TNBC细胞的生长抑制作用效果。高浓度组合15nM LBH589联合2μΜ汉防己甲素可以直接杀伤TNBC细胞诱导细胞凋亡。低浓度组合5nM LBH589联合2μΜ汉防己甲素没有显著急性细胞毒性而能协同抑制TNBC细胞干性和细胞迁移。进一步机制研究表明CDC25A表达下调而诱导细胞G1期阻滞参与调控LBH589联合汉防己甲素协同抗TNBC。本项目探索了一种新的双药联合治疗策略，进一步推动LBH589 作为抗肿瘤靶向药物的临床使用，为扩大其临床治疗肿瘤类型提供了实验依据，也为汉防己甲素发展为抗肿瘤药物提供又一证据，进一步丰富了本民族传统中药的抗肿瘤应用，给未来TNBC治疗临床试验奠定了理论基础。