Oroxylin A, a natural anticancer flavonoid compound, induces differentiation of t(8;21)-positive Kasumi-1 and primary acute myeloid leukemia cells
Oroxylin A, a natural anticancer flavonoid compound, induces differentiation of t(8;21)-positive Kasumi-1 and primary acute myeloid leukemia cells
复制标题
Oroxylin A 是一种天然抗癌类黄酮化合物，可诱导 t(8;21) 阳性 Kasumi-1 和原代急性髓系白血病细胞分化
DOI：
10.1007/s00432-016-2160-1
复制
发表时间：
2016-07-01
影响因子：
3.6
通讯作者：
Lu, Na
中科院分区：
文献类型：
--
作者：
Hui, Hui;Zhang, Xiaoxiao;Lu, Na
目的：AML 1/ETO融合基因是t（8;21）阳性急性髓系白血病（AML）的致病基因之一。Oroxylin A（OA）对其他癌细胞具有抗癌作用。本研究采用MTT法检测OA对t（8; 21）阳性的Kasumi-1细胞和原代AML细胞活性的影响。通过NBT还原试验、流式细胞术分析CD 11b/CD 14和Giemsa染色检测细胞分化。蛋白质表达通过蛋白质印迹法测定。免疫荧光法检测OA对HDAC-1表达的影响。结果：OA能显著增强AML 1/ETO阳性细胞的NBT还原活性和CD 11b/CD 14表达。Giemsa染色结果也显示OA可引起细胞形态学改变，核质比降低，提示OA诱导细胞分化。进一步的研究表明，OA在体内外均能降低AML 1/ETO融合蛋白的表达，下调HDAC-1蛋白水平。OA诱导分化相关蛋白C/EBPα和P21表达增加。治疗后组蛋白乙酰化水平也明显升高。结论：OA有可能成为AML 1/ETO阳性AML诱导分化治疗的新候选药物，其作用机制有待进一步研究。
Purpose:AML1/ETO fusion gene is one of disease-causing genes of t(8;21)-positive acute myeloid leukemia (AML). Oroxylin A (OA) has showed anticancer effects on other cancer cells. Here, studies were conducted to determine the antileukemia effect of OA on t(8;21)-positive AML cells in vitro and in vivo.Materials and methods:The effects of OA on cell viability of t(8;21)-positive Kasumi-1 and primary AML cells were analyzed by MTT assay. Cell differentiation was examined by NBT reduction assay, flow cytometry analysis for CD11b/CD14, and Giemsa stain. Protein expressions were determined by Western blots. Immunofluorescence assay was used to verify the effect of OA on HDAC-1 expression in vivo. Immunohistochemical staining was applied to evaluate leukemic infiltration of AML-bearing NOD/SCID mice.Results:OA enhanced NBT reduction activity and CD11b/CD14 expression of AML1/ETO-positive AML cells markedly. Results of Giemsa staining also demonstrated that OA could induce the morphologic changes with reduction of nuclear/cytoplasmic ratios, suggesting the cell differentiation induced by OA. Further study showed that OA decreased the expression of fusion protein AML1/ETO and down-regulated HDAC-1 protein levels in vitro and in vivo. Moreover, OA increased the expression of differentiation-related proteins C/EBPα and P21. Acetylation levels of histones were also advanced obviously after treatment of OA. In vivo study indicated that OA could prolong the survival of AML-bearing NOD/SCID mice and reduce leukocytic infiltration of the spleen.Conclusions:All these results suggested that OA might be a novel candidate agent for differentiation therapy for AML1/ETO-positive AML and the mechanism required further investigation.