Priority Medical Department, General Hospital of Central Theater Command, Wuhan, 430070, Hubei, China.
Priority Medical Department, General Hospital of Central Theater Command, Wuhan, 430070, Hubei, China.
Mol Cell Probes. 2023 Jun;69:101911. doi: 10.1016/j.mcp.2023.101911. Epub 2023 Apr 5.
Tumor necrosis factor receptor-related factor 7 (TRAF7) can regulate cell differentiation and apoptosis, but its specific functional mechanism in the pathological process of acute myeloid leukemia (AML) closely related to differentiation and apoptosis disorders is largely unclear. In this study, TRAF7 was found to be lowly expressed in AML patients and a variety of myeloid leukemia cells. TRAF7 was overexpressed in AML Molm-13 and chronic myeloid leukemia (CML) K562 cells by transfection with pcDNA3.1-TRAF7. CCK-8 assay and flow cytometry analysis showed that TRAF7 overexpression induced growth inhibition and apoptosis in K562 and Molm-13 cells. Measurements of glucose and lactate suggested that TRAF7 overexpression impaired glycolysis of K562 and Molm-13 cells. Cell cycle analysis indicated that most of K562 and Molm-13 cells were captured in G0/G1 phase by TRAF7 overexpression. PCR and western blot assay revealed that TRAF7 increased Kruppel-like factor 2 (KLF2) expression but decreased 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) expression in AML cells. KLF2 knockdown can counteract TRAF7-triggered PFKFB3 inhibition, and abolish TRAF7-mediated glycolysis inhibition and cell cycle arrest. KLF2 knockdown or PFKFB3 overexpression both can partially neutralize TRAF7-induced growth inhibition and apoptosis of K562 and Molm-13 cells. Moreover, Lv-TRAF7 decreased human CD45 cells in mouse peripheral blood in the xenograft mice established by NOD/SCID mice. Taken together, TRAF7 exerts anti-leukemia effects by impairing glycolysis and cell cycle progression of myeloid leukemia cells via modulating the KLF2-PFKFB3 axis.
肿瘤坏死因子受体相关因子 7（TRAF7）可以调节细胞分化和凋亡，但它在与分化和凋亡紊乱密切相关的急性髓系白血病（AML）病理过程中的具体功能机制在很大程度上尚不清楚。在本研究中，发现 TRAF7 在 AML 患者和各种髓系白血病细胞中低表达。通过转染 pcDNA3.1-TRAF7 使 AML Molm-13 和慢性髓系白血病（CML）K562 细胞中 TRAF7 过表达。CCK-8 测定和流式细胞术分析表明，TRAF7 过表达诱导 K562 和 Molm-13 细胞生长抑制和凋亡。葡萄糖和乳酸测量表明，TRAF7 过表达损害了 K562 和 Molm-13 细胞的糖酵解。细胞周期分析表明，TRAF7 过表达使大多数 K562 和 Molm-13 细胞被捕获在 G0/G1 期。PCR 和 Western blot 分析表明，TRAF7 增加了 AML 细胞中 Kruppel 样因子 2（KLF2）的表达，但降低了 6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 3（PFKFB3）的表达。KLF2 敲低可以抵消 TRAF7 触发的 PFKFB3 抑制，并消除 TRAF7 介导的糖酵解抑制和细胞周期阻滞。KLF2 敲低或 PFKFB3 过表达均可部分中和 TRAF7 诱导的 K562 和 Molm-13 细胞生长抑制和凋亡。此外，Lv-TRAF7 在 NOD/SCID 小鼠建立的异种移植小鼠中减少了小鼠外周血中的人 CD45 细胞。综上所述，TRAF7 通过调节 KLF2-PFKFB3 轴来破坏髓系白血病细胞的糖酵解和细胞周期进展，从而发挥抗白血病作用。
