Fu Dian, Shi Xiuquan, Yi Xiaoming, Wu Ding, He Haowei, Zhou Wenquan, Cheng Wen
Department of Urology, Jinling College of Clinical Medicine, Nanjing Medical University, No.305, Zhongshandong Road, Xuanwu District, Nanjing, Jiangsu, 210002, China.
Department of Urology, Nanjing Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, No.305, Zhongshandong Road, Xuanwu District, Nanjing, Jiangsu, 210002, China.
BMC Urol. 2024 Jul 16;24(1):147. doi: 10.1186/s12894-024-01534-4.
Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) has been confirmed to play oncogenic role in many cancers. However, the role and mechanism of IGF2BP2 in bladder cancer (BCa) still deserves to be further revealed.
The mRNA and protein levels of IGF2BP2 and neuronilin-1 (NRP1) were detected by real-time quantitative PCR (RT-qPCR) and western blot. Cell proliferation, apoptosis, migration and invasion were determined using colony formation assay, EdU assay, CCK8 assay, flow cytometry and transwell assay. Xenograft tumor model was conducted to evaluate the role of IGF2BP2 in vivo. THP-1-M0 macrophages were co-cultured with the condition medium (CM) of BCa cells to induce polarization. M2 macrophage polarization was assessed by detecting the mRNA levels of M2 macrophage markers using RT-qPCR and measuring the proportion of M2 macrophage markers using flow cytometry. Moreover, MeRIP and RIP assay were performed to assess m6A level and the interaction between IGF2BP2 and NRP1.
IGF2BP2 and NRP1 were upregulated in BCa tissues and cells. IGF2BP2 knockdown suppressed BCa cell growth and metastasis, as well as inhibited BCa tumor growth. After THP-1-M0 macrophages were co-cultured with the CM of BCa cells, the levels of M2 macrophage markers were markedly enhanced, while this effect was abolished by IGF2BP2 knockdown. IGF2BP2 level was positively correlated with NRP1 level, and it could increase NRP1 mRNA stability. NRP1 overexpression reversed the suppressive effect of IGF2BP2 knockdown on M2 macrophage polarization and BCa cell progression.
m6A-reader IGF2BP2 enhanced M2 macrophage polarization and BCa cell progression by promoting NRP1 mRNA stability.
胰岛素样生长因子2 mRNA结合蛋白2（IGF2BP2）已被证实在多种癌症中发挥致癌作用。然而，IGF2BP2在膀胱癌（BCa）中的作用及机制仍有待进一步揭示。
采用实时定量PCR（RT-qPCR）和蛋白质免疫印迹法检测IGF2BP2和神经纤毛蛋白-1（NRP1）的mRNA和蛋白水平。使用集落形成试验、EdU试验、CCK8试验、流式细胞术和Transwell试验测定细胞增殖、凋亡、迁移和侵袭。建立异种移植瘤模型以评估IGF2BP2在体内的作用。将THP-1-M0巨噬细胞与BCa细胞的条件培养基（CM）共培养以诱导极化。通过RT-qPCR检测M2巨噬细胞标志物的mRNA水平并使用流式细胞术测量M2巨噬细胞标志物的比例来评估M2巨噬细胞极化。此外，进行甲基化RNA免疫沉淀（MeRIP）和RNA免疫沉淀（RIP）试验以评估m6A水平以及IGF2BP2与NRP1之间的相互作用。
IGF2BP2和NRP1在BCa组织和细胞中上调。IGF2BP2基因敲低抑制了BCa细胞的生长和转移，并抑制了BCa肿瘤的生长。THP-1-M0巨噬细胞与BCa细胞的CM共培养后，M2巨噬细胞标志物的水平显著升高，而IGF2BP2基因敲低消除了这种作用。IGF2BP2水平与NRP1水平呈正相关，并且它可以增加NRP1 mRNA的稳定性。NRP1过表达逆转了IGF2BP2基因敲低对M2巨噬细胞极化和BCa细胞进展的抑制作用。
m6A阅读蛋白IGF2BP2通过促进NRP1 mRNA稳定性增强M2巨噬细胞极化和BCa细胞进展。
