[Effect of Dimethyl Fumarate (DMF) on T-cell Acute Lymphoblastic Leukemia].

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.富马酸二甲酯对急性T淋巴细胞白血病影响的研究.探讨富马酸二甲酯（DMF）对急性T淋巴细胞白血病的影响及潜在的作用机制，为临床治疗急性T淋巴细胞白血病提供实验基础和理论依据.探讨富马酸二甲酯（DMF）对急性T淋巴细胞白血病的影响及潜在的作用机制，为临床治疗急性T淋巴细胞白血病提供实验基础和理论依据。方法：用不同浓度DMF处理Jurkat细胞24 h，然后采用流式细胞术检测Ki67+的Jurkat细胞比例和绝对计数；同时采用Western blot方法检测DMF处理24 h之后，Jurkat细胞内核因子E2相关因子2（Nrf2）和E3泛素连接酶HACE1的蛋白水平变化；利用免疫共沉淀方法富集Jurkat细胞内Nrf2蛋白，然后采用Western blot方法检测HACE1蛋白水平；在HEK293T细胞中过表达Flag-Nrf2和不同梯度的Flag-HACE1质粒，48 h后采用Western blot检测Flag-Nrf2蛋白水平变化。结果：DMF能显著抑制Ki67+的Jurkat细胞比例和绝对计数，即DMF抑制Jurkat细胞增殖，且呈剂量依赖性（r=0.9595，r=0.9054）。DMF能显著上调Jurkat细胞内Nrf2和E3泛素连接酶HACE1蛋白水平（P<0.01，P<0.01）。Jurkat细胞内Nrf2和HACE1蛋白存在相互作用。过表达Flag-HACE1显著上调Flag-Nrf2的蛋白水平，且呈剂量依赖性（r=0.9771）.DMF能显著抑制Ki67+的Jurkat细胞比例和绝对计数，即DMF抑制Jurkat细胞增殖，且呈剂量依赖性（r=0.9595，r=0.9054）。DMF能显著上调Jurkat细胞内Nrf2和E3泛素连接酶HACE1蛋白水平（P<0.01，P<0.01）。Jurkat细胞内Nrf2和HACE1蛋白存在相互作用。过表达Flag-HACE1显著上调Flag-Nrf2的蛋白水平，且呈剂量依赖性（r=0.9771）.DMF抑制急性T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蛋白水平，HACE1与Nrf2相互作用并正向调控Nrf2蛋白水平.