Destabilization of macrophage migration inhibitory factor by 4-IPP reduces NF-κB/P-TEFb complex-mediated c-Myb transcription to suppress osteosarcoma tumourigenesis.
Destabilization of macrophage migration inhibitory factor by 4-IPP reduces NF-κB/P-TEFb complex-mediated c-Myb transcription to suppress osteosarcoma tumourigenesis.
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4-IPP 使巨噬细胞迁移抑制因子不稳定，从而减少 NF-ΔB/P-TEFb 复合物介导的 c-Myb 转录，从而抑制骨肉瘤肿瘤发生
DOI：
10.1002/ctm2.652
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发表时间：
2022-01
影响因子：
10.6
通讯作者：
Zhao F
中科院分区：
文献类型：
--
作者：
Zheng L;Feng Z;Tao S;Gao J;Lin Y;Wei X;Zheng B;Huang B;Zheng Z;Zhang X;Liu J;Shan Z;Chen Y;Chen J;Zhao F
巨噬细胞迁移抑制因子（macrophage migration inhibitory factor， MIF）作为一种炎症因子和致癌驱动蛋白，在骨肉瘤微环境中起着至关重要的作用。虽然4‐碘‐6‐苯基嘧啶（4‐IPP）可以灭活MIF的生物学功能，但其抗骨肉瘤的作用和分子机制尚未被研究。在这项研究中，我们发现MIF抑制剂4 - IPP是一种具有抗肿瘤和抗破骨细胞生成功能的骨肉瘤特异性双效应药物。通过伤口愈合实验、细胞周期分析、菌落形成实验、CCK‐8实验、细胞凋亡分析和Transwell迁移/侵袭实验来评估4‐IPP的抗癌作用。通过荧光素酶报告基因、染色质免疫沉淀试验、免疫荧光和共免疫沉淀分析，证实了NF - κB/P - TEFb复合物对c - Myb和STUB1介导的蛋白酶体依赖性MIF蛋白降解的转录调控。4‐IPP对肿瘤生长和转移的影响通过HOS衍生的尾静脉转移模型和皮下和原位异种移植肿瘤模型进行评估。在体外实验中，4‐IPP通过抑制NF‐κB通路显著降低骨肉瘤细胞的增殖和转移。4‐IPP阻碍了MIF与CD74和p65的结合。此外，4‐IPP抑制MIF阻断下游NF‐κB/P‐TEFb复合物的形成，导致c‐Myb转录下调。有趣的是，4‐IPP的实施可以通过stub1e3配体介导小分子诱导的MIF蛋白蛋白酶体降解。然而，4‐IPP仍然会中断MIF介导的骨肉瘤细胞与破骨细胞之间的通讯，从而促进破骨细胞的发生。值得注意的是，在小鼠体内模型中，4‐IPP强烈地减少了HOS来源的异种移植物骨肉瘤的肿瘤发生和转移。我们的研究结果表明，靶向MIF蛋白的小分子4 - IPP通过同时灭活MIF的生物学功能并促进其蛋白酶体降解来发挥抗骨肉瘤的作用。用4‐IPP直接破坏MIF蛋白的稳定性可能是治疗骨肉瘤的一种有前景的治疗策略。1. MIF抑制剂4‐IPP通过抑制NF‐κB通路减少骨肉瘤细胞的增殖和转移。2. MIF抑制剂4 - IPP阻断了MIF/CD74诱导的NF - κB/P - TEFb复合物介导的骨肉瘤c - Myb转录。3. MIF抑制剂4‐IPP通过stub1e3连接酶介导MIF蛋白蛋白酶体降解。
As an inflammatory factor and oncogenic driver protein, the pleiotropic cytokine macrophage migration inhibitory factor (MIF) plays a crucial role in the osteosarcoma microenvironment. Although 4‐iodo‐6‐phenylpyrimidine (4‐IPP) can inactivate MIF biological functions, its anti‐osteosarcoma effect and molecular mechanisms have not been investigated. In this study, we identified the MIF inhibitor 4‐IPP as a specific double‐effector drug for osteosarcoma with both anti‐tumour and anti‐osteoclastogenic functions. The anti‐cancer effects of 4‐IPP were evaluated by wound healing assay, cell cycle analysis, colony formation assay, CCK‐8 assay, apoptosis analysis, and Transwell migration/invasion assays. Through the application of a luciferase reporter, chromatin immunoprecipitation assays, and immunofluorescence and coimmunoprecipitation analyses, the transcriptional regulation of the NF‐κB/P‐TEFb complex on c‐Myb‐ and STUB1‐mediated proteasome‐dependent MIF protein degradation was confirmed. The effect of 4‐IPP on tumour growth and metastasis was assessed using an HOS‐derived tail vein metastasis model and subcutaneous and orthotopic xenograft tumour models. In vitro, 4‐IPP significantly reduced the proliferation and metastasis of osteosarcoma cells by suppressing the NF‐κB pathway. 4‐IPP hindered the binding between MIF and CD74 as well as p65. Moreover, 4‐IPP inhibited MIF to interrupt the formation of downstream NF‐κB/P‐TEFb complexes, leading to the down‐regulation of c‐Myb transcription. Interestingly, the implementation of 4‐IPP can mediate small molecule‐induced MIF protein proteasomal degradation via the STUB1 E3 ligand. However, 4‐IPP still interrupted MIF‐mediated communication between osteosarcoma cells and osteoclasts, thus promoting osteoclastogenesis. Remarkably, 4‐IPP strongly reduced HOS‐derived xenograft osteosarcoma tumourigenesis and metastasis in an in vivo mouse model. Our findings demonstrate that the small molecule 4‐IPP targeting the MIF protein exerts an anti‐osteosarcoma effect by simultaneously inactivating the biological functions of MIF and promoting its proteasomal degradation. Direct destabilization of the MIF protein with 4‐IPP may be a promising therapeutic strategy for treating osteosarcoma. 1. MIF inhibitor 4‐IPP reduced the proliferation and metastasis of osteosarcoma cells by suppressing the NF‐κB pathway. 2. MIF inhibitor 4‐IPP interrupted the MIF/CD74‐induced NF‐κB/P‐TEFb complex‐mediated c‐Myb transcription in osteosarcoma. 3. MIF inhibitor 4‐IPP mediated MIF protein proteasomal degradation via the STUB1 E3 ligase.
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影响因子：
8.8
作者：
Goguet-Surmenian, E.;Richard-Fiardo, P.;Guillemot, E.;Benchetrit, M.;Gomez-Brouchet, A.;Buzzo, P.;Karimdjee-Soilihi, B.;Alemanno, P.;Michiels, J-F;Schmid-Alliana, A.;Schmid-Antomarchi, H.
通讯作者：
Schmid-Antomarchi, H.
影响因子：
32.4
作者：
Bucala, Richard;Donnelly, Seamas C.
通讯作者：
Donnelly, Seamas C.
影响因子：
15.9
作者：
Lu, Jinchang;Song, Guohui;Wang, Jin
通讯作者：
Wang, Jin
影响因子：
16
作者：
Barboric, M;Nissen, RM;Peterlin, BM
通讯作者：
Peterlin, BM
影响因子：
5.2
作者：
Chao, Chia-Chia;Lee, Chiang-Wen;Liu, Ju-Fang
通讯作者：
Liu, Ju-Fang