Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, China.
Immun Inflamm Dis. 2023 Feb;11(2):e789. doi: 10.1002/iid3.789.
Diffuse large B-cell lymphoma (DLBCL) remains the most frequent subpopulation of lymphoma, and N6-methyladenosine (m6A) was implicated in the DLBCL progression. Herein, we sought to decipher the m6A-asociated mechanism of NEDD1 in DLBCL development.
The NEDD1 expression profile in DLBCL was assessed by quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot. NEDD1 was artificially downregulated or upregulated in DLBCL cells, followed by EdU, Transwell assays and flow cytometry. The Hedgehog pathway activity was assayed by a dual-luciferase assay. The m6A methylation of NEDD1 in DLBCL was assessed by meRIP-qPCR, and the regulatory mechanism of METTL3 on NEDD1 was validated. The LDH assay was conducted to examine the impact of CD8 T cells on DLBCL cells. The DLBCL cells were administrated into mice to evaluate the tumorigenic activity and ki-67 activity in tumor tissues.
NEDD1 was overexpressed in DLBCL. Depletion of NEDD1 inhibited the aggressiveness of SU-DHL-8 and OCI-LY1 cells, whereas overexpression of NEDD1 expedited the aggressiveness of SU-DHL-8 and OCI-LY1 cells. METTL3 promoted NEDD1 translation in an m6A-dependent manner via YTHDF1. Depletion of METTL3 inhibited SU-DHL-8 and OCI-LY1 cell activity through regulation of NEDD1. NEDD1 reversed the repressive effect of METTL3 loss on the aggressiveness of SU-DHL-8 and OCI-LY1 cells. NEDD1 activated the Hedgehog signaling to promote immune escape of DLBCL.
METTL3 promotes translation of NEDD1 via YTHDF1-depedndent m6A modification, thereby activating the Hedgehog signaling pathway to promote immune escape of DLBCL cells.
弥漫性大 B 细胞淋巴瘤（DLBCL）仍然是最常见的淋巴瘤亚群，N6-甲基腺苷（m6A）被认为与 DLBCL 的进展有关。在此，我们试图破译 NEDD1 在 DLBCL 发展中的 m6A 相关机制。
通过定量实时聚合酶链反应（RT-qPCR）和 Western blot 评估 DLBCL 中的 NEDD1 表达谱。在 DLBCL 细胞中人工下调或上调 NEDD1，然后进行 EdU、Transwell 测定和流式细胞术。通过双荧光素酶测定法测定 Hedgehog 途径活性。通过 meRIP-qPCR 评估 DLBCL 中 NEDD1 的 m6A 甲基化，并验证 METTL3 对 NEDD1 的调节机制。LDH 测定法用于检查 CD8 T 细胞对 DLBCL 细胞的影响。将 DLBCL 细胞施用于小鼠以评估肿瘤组织中的肿瘤发生活性和 ki-67 活性。
NEDD1 在 DLBCL 中过表达。NEDD1 的缺失抑制了 SU-DHL-8 和 OCI-LY1 细胞的侵袭性，而 NEDD1 的过表达则加速了 SU-DHL-8 和 OCI-LY1 细胞的侵袭性。METTL3 通过 YTHDF1 以 m6A 依赖性方式促进 NEDD1 的翻译。METTL3 的缺失通过调节 NEDD1 抑制了 SU-DHL-8 和 OCI-LY1 细胞的活性。NEDD1 逆转了 METTL3 缺失对 SU-DHL-8 和 OCI-LY1 细胞侵袭性的抑制作用。NEDD1 激活 Hedgehog 信号通路促进 DLBCL 细胞的免疫逃逸。
METTL3 通过 YTHDF1 依赖性 m6A 修饰促进 NEDD1 的翻译，从而激活 Hedgehog 信号通路促进 DLBCL 细胞的免疫逃逸。
