The First Affiliated Hospital, Department of Blood Transfusion, Hengyang Medical School, University of South China, Hengyang, 421001, Hunan Province, People's Republic of China.
The First Affiliated Hospital, Department of Laboratory Medicine, Hengyang Medical School, University of South China, Hengyang, 421001, Hunan Province, People's Republic of China.
Ann Hematol. 2023 Jul;102(7):1801-1810. doi: 10.1007/s00277-023-05283-6. Epub 2023 May 24.
N6-methyladenosine (m6A) methyltransferase-like 3 (METTL3) has been confirmed to be involved in multiple myeloma (MM) progression, and basic leucine zipper and W2 domains 2 (BZW2) is considered to be a regulator for MM development. However, whether METTL3 mediates MM progression by regulating BZW2 remains unclear. The messenger RNA (mRNA) and protein levels of METTL3 and BZW2 in MM specimens and cells were determined using quantitative real-time PCR and western blot analysis. Cell proliferation and apoptosis were assessed by cell counting kit 8 assay, 5-ethynyl-2'-deoxyuridine assay, colony formation assay, and flow cytometry. Methylated RNA immunoprecipitation-qPCR was used to detect the m6A modification level of BZW2. Xenograft tumor models were constructed to confirm the effect of METTL3 knockdown on MM tumor growth in vivo. Our results showed that BZW2 was upregulated in MM bone marrow specimens and cells. BZW2 downregulation reduced MM cell proliferation and promoted apoptosis, while its overexpression enhanced MM cell proliferation and inhibited apoptosis. METTL3 was highly expressed in MM bone marrow specimens, and its expression was positively correlated with BZW2 expression. BZW2 expression was positively regulated by METTL3. Mechanistically, METTL3 could upregulate BZW2 expression by modulating its m6A modification. Additionally, METTL3 accelerated MM cell proliferation and restrained apoptosis via increasing BZW2 expression. In vivo experiments showed that METTL3 knockdown reduced MM tumor growth by decreasing BZW2 expression. In conclusion, these data indicated that METTL3-mediated the m6A methylation of BZW2 to promote MM progression, suggesting a novel therapeutic target for MM.
N6-甲基腺苷（m6A）甲基转移酶样 3（METTL3）已被证实参与多发性骨髓瘤（MM）的进展，而碱性亮氨酸拉链和 W2 结构域 2（BZW2）被认为是 MM 发育的调节剂。然而，METTL3 是否通过调节 BZW2 来介导 MM 的进展尚不清楚。采用实时定量 PCR 和 Western blot 分析检测 MM 标本和细胞中 METTL3 和 BZW2 的 mRNA 和蛋白水平。通过细胞计数试剂盒 8 检测、5-乙炔基-2'-脱氧尿苷检测、集落形成检测和流式细胞术评估细胞增殖和凋亡。采用甲基化 RNA 免疫沉淀-qPCR 检测 BZW2 的 m6A 修饰水平。构建异种移植肿瘤模型以证实 METTL3 敲低对 MM 肿瘤在体内生长的影响。结果显示，BZW2 在 MM 骨髓标本和细胞中上调。BZW2 下调减少 MM 细胞增殖并促进凋亡，而过表达增强 MM 细胞增殖并抑制凋亡。METTL3 在 MM 骨髓标本中高表达，其表达与 BZW2 表达呈正相关。BZW2 的表达受 METTL3 的正向调节。机制上，METTL3 通过调节 BZW2 的 m6A 修饰来上调 BZW2 的表达。此外，METTL3 通过增加 BZW2 的表达来加速 MM 细胞的增殖并抑制凋亡。体内实验表明，METTL3 敲低通过降低 BZW2 的表达来减少 MM 肿瘤的生长。总之，这些数据表明 METTL3 通过介导 BZW2 的 m6A 甲基化促进 MM 的进展，提示 MM 的一个新的治疗靶点。
