Center of Medical Genetics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Hualien 97004, Taiwan.
Department of Molecular Biology and Human Genetics, Tzu Chi University, Hualien 97004, Taiwan.
Int J Mol Sci. 2022 Jul 2;23(13):7392. doi: 10.3390/ijms23137392.
Acute myeloid leukemia (AML) is characterized by the dysregulation of hematopoietic cell proliferation, resulting in the accumulation of immature myeloid cells in bone marrow. 5-Demethylnobiletin (5-demethyl NOB), a citrus 5-hydroxylated polymethoxyflavone, has been reported to exhibit various bioactivities, such as antioxidant, anti-inflammatory and anticancer properties. In this study, we investigated the antileukemic effects of 5-demethyl NOB and its underlying molecular mechanisms in human AML cells. We found that 5-demethyl NOB (20−80 μM) significantly reduced human leukemia cell viability, and the following trend of effectiveness was observed: THP-1 ≈ U-937 > HEL > HL-60 > K562 cells. 5-Demethyl NOB (20 and 40 μM) modulated the cell cycle through the regulation of p21, cyclin E1 and cyclin A1 expression and induced S phase arrest. 5-Demethyl NOB also promoted leukemia cell apoptosis and differentiation. Microarray-based transcriptome, Gene Ontology (GO) and Gene Set Enrichment Analysis (GSEA) of differentially expressed genes (DEGs) analysis showed that the expression of inhibitor of differentiation/DNA binding 1 (ID1), a gene associated with the GO biological process (BP) cell population proliferation (GO: 0008283), was most strongly suppressed by 5-demethyl NOB (40 μM) in THP-1 cells. We further demonstrated that 5-demethyl NOB-induced ID1 reduction was associated with the inhibition of leukemia cell growth. Moreover, DEGs involved in the hallmark gene set NF-κB/TNF-α signaling pathway were markedly enriched and downregulated by 5-demethyl NOB. Finally, we demonstrated that 5-demethyl NOB (20 and 40 μM), combined with cytarabine, synergistically reduced THP-1 and U-937 cell viability. Our current findings support that 5-demethyl NOB dramatically suppresses leukemia cell proliferation and may serve as a potential phytochemical for human AML chemotherapy.
急性髓细胞白血病（AML）的特征是造血细胞增殖失调，导致骨髓中未成熟髓细胞的积累。5-去甲基诺比汀（5-demethyl NOB），一种柑橘类 5-羟基化多甲氧基黄酮，已被报道具有多种生物活性，如抗氧化、抗炎和抗癌特性。在本研究中，我们研究了 5-去甲基诺比汀在人 AML 细胞中的抗白血病作用及其潜在的分子机制。我们发现，5-去甲基诺比汀（20-80μM）显著降低人白血病细胞活力，其效果呈以下趋势：THP-1≈U-937＞ HEL＞HL-60＞K562 细胞。5-去甲基诺比汀（20 和 40μM）通过调节 p21、细胞周期蛋白 E1 和细胞周期蛋白 A1 的表达来调节细胞周期，并诱导 S 期阻滞。5-去甲基诺比汀还促进白血病细胞凋亡和分化。基于微阵列的转录组、基因本体论（GO）和基因集富集分析（GSEA）的差异表达基因（DEGs）分析表明，分化/DNA 结合抑制因子 1（ID1）的表达受到 5-去甲基诺比汀（40μM）的强烈抑制，ID1 是与 GO 生物过程（BP）细胞群体增殖（GO:0008283）相关的基因。我们进一步证明，5-去甲基诺比汀诱导的 ID1 减少与白血病细胞生长的抑制有关。此外，5-去甲基诺比汀显著富集并下调了参与 NF-κB/TNF-α 信号通路标志性基因集的 DEGs。最后，我们证明，5-去甲基诺比汀（20 和 40μM）与阿糖胞苷联合使用可协同降低 THP-1 和 U-937 细胞活力。我们目前的研究结果支持 5-去甲基诺比汀可显著抑制白血病细胞增殖，可能成为人类 AML 化疗的潜在植物化学物质。
