Central Laboratory of Yong-Chuan Hospital, Chongqing Medical University, Chongqing 402160, China.
Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
Cell Signal. 2023 May;105:110626. doi: 10.1016/j.cellsig.2023.110626. Epub 2023 Feb 7.
Studies have confirmed that acute myeloid leukemia (AML) cells with DNA methyltransferase 3A Arg882His (DNMT3A R882H) mutation show an increased proliferation capability. However, the associated mechanism is still unclear. Glycolysis is involved in regulating malignant proliferation of cancer cell. Hence, we analyzed whether the DNMT3A R882H mutation interferes with glycolysis and thereby influences AML cell proliferation.
We generated AML cell line carrying a DNMT3A-R882H mutation and compared it with the wild type (DNMT3A-WT) with regard to glycolysis regulation. Moreover, we analyzed the cell line's proliferation and apoptosis by a CCK-8 assay, western blotting, and flow cytometry. The role of NRF2/NQO1 signaling in regulating glycolysis was investigated by NRF2-knockdown and Brusatol (specific inhibitor of NRF2) treatment.
DNMT3A R882H cells had a higher glucose transport capacity compared to WT cells and their viability could be reduced by glucose deprivation. Moreover, daunorubicin had a slight inhibitory effect on glycolysis while glycolysis inhibition re-sensitized mutant cells to daunorubicin. Obviously, DNMT3A R882H mutation activated the NRF2/NQO1 pathway and enhanced the glycolytic activity in mutant cells.
Taken together, these results suggest a novel mechanism by which a DNMT3A R882H mutation promotes glycolysis via activation of NRF2/NQO1 pathway. A parallel glycolysis inhibition adds to the anticancer effects of daunorubicin which might lead to a novel therapeutic approach for the treatment of AML patients carrying a DNMT3A R882H mutation.
研究证实，具有 DNA 甲基转移酶 3A Arg882His（DNMT3A R882H）突变的急性髓系白血病（AML）细胞表现出更高的增殖能力。然而，相关机制尚不清楚。糖酵解参与调节癌细胞的恶性增殖。因此，我们分析了 DNMT3A R882H 突变是否干扰糖酵解从而影响 AML 细胞增殖。
我们构建了携带 DNMT3A-R882H 突变的 AML 细胞系，并与野生型（DNMT3A-WT）进行了比较，以研究其对糖酵解的调节作用。此外，我们通过 CCK-8 测定、Western blot 和流式细胞术分析了细胞系的增殖和凋亡情况。通过 NRF2 敲低和 Brusatol（NRF2 的特异性抑制剂）处理，研究了 NRF2/NQO1 信号通路在调节糖酵解中的作用。
与 WT 细胞相比，DNMT3A R882H 细胞具有更高的葡萄糖转运能力，且葡萄糖剥夺可降低其活力。此外，柔红霉素对糖酵解仅有轻微的抑制作用，而抑制糖酵解则可使突变细胞对柔红霉素重新敏感。显然，DNMT3A R882H 突变激活了 NRF2/NQO1 通路，并增强了突变细胞的糖酵解活性。
综上所述，这些结果表明，DNMT3A R882H 突变通过激活 NRF2/NQO1 通路促进糖酵解，这为其促进肿瘤发生提供了一种新的机制。并行的糖酵解抑制作用增强了柔红霉素的抗癌作用，这可能为携带 DNMT3A R882H 突变的 AML 患者的治疗提供一种新的治疗方法。
