天然HDAC-1/8抑制剂黄芩素对核心结合因子急性髓系白血病具有治疗作用。
Natural HDAC-1/8 inhibitor baicalein exerts therapeutic effect in CBF-AML.
作者信息
Yu Xiaoxuan, Li Hui, Hu Po, Qing Yingjie, Wang Xiangyuan, Zhu Mengyuan, Wang Hongzheng, Wang Zhanyu, Xu Jingyan, Guo Qinglong, Hui Hui
机构信息
State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, Jiangsu Key Laboratory of Drug Design and Optimization, China Pharmaceutical University, China Pharmaceutical University, Nanjing, Jiangsu, China.
Department of Pharmacology, School of medicine & Holostic integrative medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China.
出版信息
Clin Transl Med. 2020 Aug;10(4):e154. doi: 10.1002/ctm2.154.
BACKGROUND
Although targeting histone deacetylases (HDACs) may be an effective strategy for core binding factor-acute myeloid leukemia (CBF-AML) harboring t(8;21) or inv(16), HDAC inhibitors are reported to be limited by drug-resistant characteristic. Our purpose is to evaluate the anti-leukemia effects of Baicalein on CBF-AML and clarify its underlying mechanism.
METHODS
Enzyme activity assay was used to measure the activity inhibition of HDACs. Rhodamine123 and RT-qPCR were employed to evaluate the distribution of drugs and the change of ATP-binding cassette (ABC) transporter genes. CCK8, Annexin V/PI, and FACS staining certified the effects of Baicalein on cell growth, apoptosis, and differentiation. Duolink and IP assay assessed the interaction between HDAC-1 and ubiquitin, HSP90 and AML1-ETO, and Ac-p53 and CBFβ-MYH11. AML cell lines and primary AML cells-bearing NOD/SCID mice models were used to evaluate the anti-leukemic efficiency and potential mechanism of Baicalein in vivo.
RESULTS
Baicalein showed HDAC-1/8 inhibition to trigger growth suppression and differentiation induction of AML cell lines and primary AML cells. Although the inhibitory action on HDAC-1 was mild, Baicalein could induce the degradation of HDAC-1 via ubiquitin proteasome pathway, thereby upregulating the acetylation of Histone H3 without promoting ABC transporter genes expression. Meanwhile, Baicalein increased the acetylation of HSP90 and lessened its connection to AML1/ETO, consequently leading to degradation of AML1-ETO in t(8;21)q(22;22) AML cells. In inv(16) AML cells, Baicalein possessed the capacity of apoptosis induction accompanied with p53-mediated apoptosis genes expression. Moreover, CBFβ-MYH11-bound p53 acetylation was restored via HDAC-8 inhibition induced by Baicalein contributing the diminishing of survival of CD34 inv(16) AML cells.
CONCLUSIONS
These findings improved the understanding of the epigenetic regulation of Baicalein, and warrant therapeutic potential of Baicalein for CBF-AML.
背景
尽管靶向组蛋白去乙酰化酶（HDACs）可能是治疗携带t(8;21)或inv(16)的核心结合因子急性髓系白血病（CBF-AML）的有效策略，但据报道HDAC抑制剂受耐药特性限制。我们的目的是评估黄芩苷对CBF-AML的抗白血病作用并阐明其潜在机制。
方法
采用酶活性测定法检测HDACs的活性抑制情况。使用罗丹明123和RT-qPCR评估药物分布及ATP结合盒（ABC）转运蛋白基因的变化。CCK8、Annexin V/PI和流式细胞术染色验证黄芩苷对细胞生长、凋亡和分化的影响。Duolink和免疫沉淀试验评估HDAC-1与泛素、HSP90与AML1-ETO以及乙酰化p53与CBFβ-MYH11之间的相互作用。使用AML细胞系和携带原发性AML细胞的NOD/SCID小鼠模型评估黄芩苷在体内的抗白血病效率和潜在机制。
结果
黄芩苷显示出对HDAC-1/8的抑制作用，从而引发AML细胞系和原发性AML细胞的生长抑制和分化诱导。尽管对HDAC-1的抑制作用较弱，但黄芩苷可通过泛素蛋白酶体途径诱导HDAC-1降解，从而上调组蛋白H3的乙酰化水平，且不促进ABC转运蛋白基因表达。同时，黄芩苷增加了HSP90的乙酰化水平并减少其与AML1/ETO的连接，从而导致t(8;21)q(22;22) AML细胞中AML1-ETO的降解。在inv(16) AML细胞中，黄芩苷具有诱导凋亡的能力，并伴有p53介导的凋亡基因表达。此外，黄芩苷诱导的HDAC-8抑制恢复了CBFβ-MYH11结合的p53乙酰化，从而降低了CD34+ inv(16) AML细胞的存活率。
结论
这些发现增进了对黄芩苷表观遗传调控的理解，并证明了黄芩苷对CBF-AML的治疗潜力。
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