Liu Jing, Lv Na, Zhou Lei, Li Yan, Yu Li
Clinics of Cadre, Department of Outpatient, First Medical Center of Chinese PLA General Hospital, Beijing 100853, China.
Department of Hematology, Carson International Cancer Center, School of Medicine, Shenzhen University, Shenzhen 518060, China.
Transl Cancer Res. 2020 Feb;9(2):827-839. doi: 10.21037/tcr.2019.12.07.
t(8;21) acute myeloid leukemia (AML) is a highly heterogenous hematological malignancy. Histone deacetylases inhibitors (HDACi) are a group of small-molecule compounds with extensive anti-tumor activity. Chidamide (CS055) is a selective HDACi independently developed by China. We aimed to investigate its anti-tumor activity and mechanism in t(8;21) AML cells.
Human AML SKNO-1 cells were transfected with AML1/ETO-siRNA to obtain SKNO-1-si-A/E cells. Human acute monocytic leukemia U937 cells were transfected with AML1/ETO fusion gene to obtain U937-A/E cells. AML1/ETO-positive (Kasumi-1, U937-A/E and SKNO-1) and AML1/ETO-negative AML cells (HL-60, U937, SKNO-1-siA/E) were treated with chidamide (0.125, 0.25 and 0.5 µM). Cell proliferation was evaluated by CCK-8 assay. Cell apoptosis and cell cycle was detected by flow cytometry. Microarray profiling of SKNO-1 cells was performed. The level of histone 3 acetylation and ERK1/2 phosphorylation was determined by Western blot. AML1/ETO and C-KIT mRNA expression was detected by real time quantitative PCR (RT-qPCR). Kasumi-1 and SKNO-1 cells were treated with U0126, a special inhibitor of ERK1/2, and cell proliferation and ERK1/2 phosphorylation level was examined.
Chidamide inhibited the proliferation, induced cell cycle arrest, and stimulated cell apoptosis of AML1/ETO-positive AML cells. Microarray profiling showed that 13 differentially expressed genes were involved both in the "ERK1/2" pathway and "apoptosis" functions. Chidamide inhibited histone 3 acetylation and ERK1/2 phosphorylation in AML1/ETO-positive AML cells. U0126 inhibited the proliferation of AML1/ETO-positive AML cells via regulating the ERK1/2 pathway.
Our results suggested that chidamide inhibits t(8;21) AML cell proliferation and AMK1/ETO and C-KIT expression by inhibiting ERK1/2 signaling pathway.
t(8;21)急性髓系白血病（AML）是一种高度异质性的血液系统恶性肿瘤。组蛋白去乙酰化酶抑制剂（HDACi）是一类具有广泛抗肿瘤活性的小分子化合物。西达本胺（CS055）是中国自主研发的一种选择性HDACi。我们旨在研究其对t(8;21) AML细胞的抗肿瘤活性及机制。
用AML1/ETO小干扰RNA转染人AML SKNO-1细胞以获得SKNO-1-si-A/E细胞。用AML1/ETO融合基因转染人急性单核细胞白血病U937细胞以获得U937-A/E细胞。用西达本胺（0.125、0.25和0.5 μM）处理AML1/ETO阳性（Kasumi-1、U937-A/E和SKNO-1）及AML1/ETO阴性AML细胞（HL-60、U937、SKNO-1-siA/E）。通过CCK-8法评估细胞增殖。通过流式细胞术检测细胞凋亡和细胞周期。对SKNO-1细胞进行基因芯片分析。通过蛋白质印迹法测定组蛋白3乙酰化水平和ERK1/2磷酸化水平。通过实时定量PCR（RT-qPCR）检测AML1/ETO和C-KIT mRNA表达。用ERK1/2特异性抑制剂U0126处理Kasumi-1和SKNO-1细胞，并检测细胞增殖和ERK1/2磷酸化水平。
西达本胺抑制AML1/ETO阳性AML细胞的增殖，诱导细胞周期阻滞，并刺激细胞凋亡。基因芯片分析显示，13个差异表达基因同时参与“ERK1/2”通路和“凋亡”功能。西达本胺抑制AML1/ETO阳性AML细胞中的组蛋白3乙酰化和ERK1/2磷酸化。U0126通过调节ERK1/2通路抑制AML1/ETO阳性AML细胞的增殖。
我们的结果表明，西达本胺通过抑制ERK1/2信号通路抑制t(8;21) AML细胞增殖以及AMK1/ETO和C-KIT表达。
