Xu Jin-Ge, Cheng Qiao, Zhang Gui-Hua, Kong Li-Ping, Li Li, Liu Kai-Ge, Wu Jin-Yan, Zhang Qiu-Rong
Department of Hematology, The Second Affiliated Hospital of Xuzhou Medical University, General Hospital of Xuzhou Coal Mining Group, Xuzhou 221006, Jiangsu Province, China,E-mail:
Department of Hematology, The First Affiliated Hospital of Soochow University, Suzhou 215000, Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2022 Feb;30(1):1-5. doi: 10.19746/j.cnki.issn.1009-2137.2022.01.01.
To explore the effect and possible mechanism of dimethyl fumarate (DMF) on T-cell acute lymphoblastic leukemia (T-ALL), and provide experimental and theoretical basis for the clinical treatment of T-ALL.
Jurkat cells were treated with different concentrations of DMF for 24 hours, and then the proportion and absolute count of Ki67-positive Jurkat cells were analyzed by flow cytometry. Meanwhile, the protein levels of nuclear factor-erythroid 2-related factor 2 (Nrf2) and E3 ubiquitin ligase HACE1 in Jurkat cells treated with DMF for 24 hours were evaluated by Western blot. Nrf2 proteins were co-immunoprecipitated in Jurkat cells, and then HACE1 protein was assessed by Western blot. Plasmids of Flag-Nrf2 and different gradients of Flag-HACE1 were transfected into HEK293T cells, and the levels of Flag-Nrf2 were detected by Western blot after 48 hours.
DMF could significantly inhibit the proportion and absolute count of Ki67-positive Jurkat cells, and DMF inhibited the proliferation of Jurkat cells in a dose-dependent manner (r=0.9595, r=0.9054). DMF could significantly up-regulate the protein levels of Nrf2 and E3 ubiquitin ligase HACE1 in Jurkat cells (P<0.01, P<0.01). HACE1 physically interacted with Nrf2 in Jurkat cells. Overexpression of Flag-HACE1 significantly increased the protein level of Flag-Nrf2 in a dose-dependent manner (r=0.9771).
DMF inhibits the proliferation of T-cell acute lymphoblastic leukemia cell. The mechanism may be that, DMF significantly up-regulates the protein levels of Nrf2 and E3 ubiquitin ligase HACE1, and HACE1 interacts with Nrf2 and positively regulates Nrf2 protein level.
探讨富马酸二甲酯（DMF）对T细胞急性淋巴细胞白血病（T-ALL）的作用及可能机制，为T-ALL的临床治疗提供实验和理论依据。
用不同浓度的DMF处理Jurkat细胞24小时，然后通过流式细胞术分析Ki67阳性Jurkat细胞的比例和绝对计数。同时，用蛋白质免疫印迹法评估DMF处理24小时的Jurkat细胞中核因子红系2相关因子2（Nrf2）和E3泛素连接酶HACE1的蛋白水平。在Jurkat细胞中对Nrf2蛋白进行免疫共沉淀，然后用蛋白质免疫印迹法评估HACE1蛋白。将Flag-Nrf2质粒和不同梯度的Flag-HACE1质粒转染到HEK293T细胞中，48小时后用蛋白质免疫印迹法检测Flag-Nrf2的水平。
DMF可显著抑制Ki67阳性Jurkat细胞的比例和绝对计数，且DMF以剂量依赖方式抑制Jurkat细胞的增殖（r=0.9595，r=0.9054）。DMF可显著上调Jurkat细胞中Nrf2和E3泛素连接酶HACE1的蛋白水平（P<0.01，P<0.01）。HACE1在Jurkat细胞中与Nrf2发生物理相互作用。Flag-HACE1的过表达以剂量依赖方式显著增加Flag-Nrf2的蛋白水平（r=0.9771）。
DMF抑制T细胞急性淋巴细胞白血病细胞的增殖。其机制可能是，DMF显著上调Nrf2和E3泛素连接酶HACE1的蛋白水平，且HACE1与Nrf2相互作用并正向调节Nrf2蛋白水平。
