Cinobufagin inhibits proliferation of acute myeloid leukaemia cells by repressing c-Myc pathway-associated genes
Cinobufagin inhibits proliferation of acute myeloid leukaemia cells by repressing c-Myc pathway-associated genes
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DOI：
10.1016/j.cbi.2022.109936
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发表时间：
2022-04-19
影响因子：
5.1
通讯作者：
Kaneda, Atsushi
中科院分区：
文献类型：
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作者：
Hirasaki, Yoshiro;Okabe, Atsushi;Kaneda, Atsushi
华蟾素是一种由亚洲蟾蜍(Bufo Gargarizans)分泌的心脏毒性蟾毒内酯类固醇。Bufan内酯抑制Na+/K+ATPase，具有与心脏苷类类似的作用，如从有毒草药中提取的洋地黄毒素或哇巴因。近年来，蟾酥内酯的抗癌作用已引起人们的重视，但其潜在的分子机制尚不清楚。我们选择华蟾素作为候选抗白血病药物，对华蟾素对人急性髓系白血病(AML)细胞株HL60和Kasumi-1的作用进行了转录转录分析。流式细胞仪分析显示，华蟾素可诱导两种细胞发生凋亡。经华蟾素处理的两种细胞株的RNA测序(RNA-seq)显示，根据基因本体论(GO)分析，通常下调的基因有丰富的“Myc活性途径”。华蟾素下调基因的基因集浓缩分析(GSEA)也显示两种细胞系的“MYC_TARGET_V2”具有最高的归一化浓缩分数(NES)。相反，“TNFA_Signating_Via_NFKB”、“Tfg_Signal”和“TGF_beta_Signal”等特征随着基因的上调而显著丰富。利用染色质免疫沉淀和测序的表观遗传学分析(CHIP-SEQ)证实，编码细胞死亡相关信号分子的基因随着H3K27ac的获得而上调，而c-Myc相关基因的下调并不伴随H3K27ac的改变。华蟾素是一种抗增殖的天然化合物，对急性髓系白血病具有c-Myc抑制和表观遗传调节活性。
Cinobufagin is a cardiotoxic bufanolide steroid secreted by the Asiatic toad, Bufo gargarizans. Bufanolides inhibit Na+/K+ ATPase and have similar effects as cardiac glycosides, such as digitoxin or ouabain derived from toxic herbs. Recently, the anti-cancer effects of bufanolides have gained attention, however the underlying molecular mechanisms remain unclear. Selecting cinobufagin as a candidate anti-leukaemia agent, we here conducted transcriptomic analyses on the effect of cinobufagin on human acute myeloid leukaemia (AML) cell lines, HL60 and Kasumi-1. Flow cytometry analysis showed that cinobufagin induced apoptosis in both cell lines. RNAsequencing (RNA-seq) of the two cell lines treated with cinobufagin revealed commonly downregulated genes with enrichment in the term "Myc active pathway" according to Gene Ontology (GO) analysis. Gene Set Enrichment Analysis (GSEA) of genes downregulated by cinobufagin also showed "MYC_TARGETS_V2" with the highest normalised enrichment score (NES) in both cell lines. In contrast, hallmarks such as "TNFA_SIGNALING_VIA_NFKB", "APOPTOSIS", and "TGF_BETA_SIGNALING" were significantly enriched as upregulated gene sets. Epigenetic analysis using chromatin immunoprecipitation and sequencing (ChIP-seq) confirmed that genes encoding cell death-related signalling molecules were upregulated by gain of H3K27ac, whereas downregulation of c-Myc-related genes was not accompanied by H3K27ac alteration. Cinobufagin is an anti-proliferative natural compound with c-Myc-inhibiting and epigenetic-modulating activity in acute myeloid leukaemia.