Department of Hematology, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou, China.
Department of Hematology, Fujian Institute of Hematology, Fujian Provincial Key Laboratory of Hematology, Fujian Medical University Union Hospital, Fuzhou, China.
J Cell Mol Med. 2021 Mar;25(5):2365-2376. doi: 10.1111/jcmm.15896. Epub 2021 Feb 10.
T cell acute lymphoblastic leukaemia (T-ALL) is a highly aggressive haematological cancer of the bone marrow. The abnormal expression of microRNAs (miRNAs) is reportedly involved in T-ALL development and progression. Thus, we aimed to decipher the involvement of miR-204 silencing mediated by DNA methylation in the occurrence of T cell acute lymphoblastic leukaemia (T-ALL). miR-204 expression was determined in bone marrow and peripheral blood samples from T-ALL patients by real-time quantitative PCR (RT-qPCR) with its effect on cell proliferation evaluated by functional assays. In addition, bisulphite sequencing PCR was employed to detect the DNA methylation level of the miR-204 promoter region, and the binding site between miR-204 and IRAK1 was detected by luciferase assay. We found that miR-204 was down-regulated in T cells of T-ALL patients, which was caused by the increased DNA methylation in the promoter region of miR-204. Moreover, overexpression of miR-204 inhibited T-ALL cell proliferation while enhancing their apoptosis through interleukin receptor-associated kinase 1 (IRAK1), which enhanced the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 through activation of p-p65. Thus, miR-204 modulated MMP-2 and MMP-9 through IRAK1/NF-κB signalling pathway, which was confirmed by in vivo assay. Taken together, DNA methylation-mediated miR-204 silencing increased the transcription of IRAK1, thus activating the NF-κB signalling pathway and up-regulating the downstream targets MMP-2/MMP-9.
T 细胞急性淋巴细胞白血病(T-ALL)是一种骨髓中高度侵袭性的血液系统恶性肿瘤。据报道,微小 RNA(miRNA)的异常表达参与了 T-ALL 的发生和发展。因此,我们旨在揭示 DNA 甲基化介导的 miR-204 沉默在 T 细胞急性淋巴细胞白血病(T-ALL)发生中的作用。通过实时定量 PCR(RT-qPCR)检测 T-ALL 患者骨髓和外周血样本中的 miR-204 表达,并通过功能测定评估其对细胞增殖的影响。此外,采用亚硫酸氢盐测序 PCR 检测 miR-204 启动子区域的 DNA 甲基化水平,通过荧光素酶测定检测 miR-204 与 IRAK1 之间的结合位点。我们发现,miR-204 在 T-ALL 患者的 T 细胞中下调,这是由于 miR-204 启动子区域的 DNA 甲基化增加所致。此外,miR-204 的过表达通过白细胞介素受体相关激酶 1(IRAK1)抑制 T-ALL 细胞增殖,同时通过 IRAK1/NF-κB 信号通路增强细胞凋亡,从而增强基质金属蛋白酶-2(MMP-2)和 MMP-9 的表达。因此,miR-204 通过 IRAK1/NF-κB 信号通路调节 MMP-2 和 MMP-9,这在体内实验中得到了验证。总之,DNA 甲基化介导的 miR-204 沉默增加了 IRAK1 的转录,从而激活了 NF-κB 信号通路并上调了下游靶标 MMP-2/MMP-9。