Inhibition of colorectal cancer aggressiveness by Oleanolic acid through Nur77 degradation.

IF 6.7 1区医学 Q1 CHEMISTRY, MEDICINAL Phytomedicine Pub Date : 2024-10-29 DOI:10.1016/j.phymed.2024.156192

Shan Deng, Yuping Liu, Xiyu Liu, Jialin Yu, Yan Chen, Jiege Huo

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Abstract

Background: Colorectal cancer (CRC) is the second primary malignancy in China with tough treatment challenge. Although Oleanolic acid (OA) protects against various cancers, its mechanisms in CRC are not well defined. Our previously study showed that Nur77 has CRC promoting effect. Thus, we investigated the roles of OA as Nur77 ligand and the regulatory effects on Nur77 degradation in CRC progression.

Methods: The proliferative and metastatic phenotypes of OA was examined by CCK-8, EdU, organoid culture, would healing and transwell assays, respectively. Epithelial-mesenchymal transition (EMT) properties were assessed by Western blotting (WB). The interaction between OA and Nur77 was monitored by molecular docking and Molecular Dynamics stimulation (MD). Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene set enrichment analysis (GSEA) were employed to screen the downstream regulatory pathways. The half-time and proteasome degradation of Nur77 were treated with cycloheximide (CHX) and MG132. Co-immunoprecipitation (Co-IP) and ubiquitination assays were employed to detect direct association between Nur77 and PPARγ. Rescued experiments were performed by Nur77 agonist Cytosporone B (Csn-B) treatment. The findings were verified in xenograft and in situ models.

Results: For the first time, we found the effect of OA on ubiquitination degradation. OA inhibited CRC cell survival and EMT phenotypes by suppressing Nur77. Mechanistically, OA directly bind to Nur77 and facilitated the ubiquitin degradation of Nur77. During this process, PPARγ acted as the ubiquitination activator via interacting with Nur77. Rescued experiments revealed that OA-induced inhibition was recovered by replenishing Nur77. In both subcutaneous and orthotopic CRC models, OA exhibited significant anti-tumor effect together with Nur77 inhibition.

Conclusion: We revealed a new regulatory effect of OA in CRC tumorigenesis via PPARγ-mediated Nur77 ubiquitin degradation.

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齐墩果酸通过降解 Nur77 抑制结直肠癌的侵袭性

背景结直肠癌（CRC）是中国第二大原发性恶性肿瘤，治疗难度很大。虽然齐墩果酸（OA）可预防多种癌症，但其在 CRC 中的作用机制尚未明确。我们之前的研究表明，Nur77 有促进 CRC 的作用。因此，我们研究了OA作为Nur77配体在CRC进展中的作用以及对Nur77降解的调控作用：方法：OA的增殖和转移表型分别通过CCK-8、EdU、类器官培养、愈合和透孔实验进行检测。上皮-间质转化（EMT）特性通过Western印迹（WB）进行评估。通过分子对接和分子动力学刺激（MD）监测了OA和Nur77之间的相互作用。采用京都基因组百科全书（KEGG）和基因组富集分析（GSEA）筛选下游调控通路。用环己亚胺（CHX）和 MG132 处理 Nur77 的半衰期和蛋白酶体降解。采用共免疫沉淀（Co-IP）和泛素化试验检测 Nur77 与 PPARγ 的直接关联。通过Nur77激动剂Cytosporone B (Csn-B)处理进行了挽救实验。这些发现在异种移植和原位模型中得到了验证：结果：我们首次发现了 OA 对泛素化降解的影响。结果：我们首次发现了OA对泛素化降解的影响。OA通过抑制Nur77抑制了CRC细胞的存活和EMT表型。从机制上讲，OA直接与Nur77结合并促进Nur77的泛素降解。在这一过程中，PPARγ通过与Nur77相互作用发挥泛素化激活剂的作用。挽救实验显示，OA诱导的抑制作用可通过补充Nur77而恢复。在皮下和正位 CRC 模型中，OA 在抑制 Nur77 的同时还表现出显著的抗肿瘤作用：结论：我们揭示了 OA 通过 PPARγ 介导的 Nur77 泛素降解对 CRC 肿瘤发生的新调控作用。

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来源期刊

Phytomedicine 医学-药学

CiteScore

10.30

自引率

5.10%

发文量

670

审稿时长

91 days

期刊介绍： Phytomedicine is a therapy-oriented journal that publishes innovative studies on the efficacy, safety, quality, and mechanisms of action of specified plant extracts, phytopharmaceuticals, and their isolated constituents. This includes clinical, pharmacological, pharmacokinetic, and toxicological studies of herbal medicinal products, preparations, and purified compounds with defined and consistent quality, ensuring reproducible pharmacological activity. Founded in 1994, Phytomedicine aims to focus and stimulate research in this field and establish internationally accepted scientific standards for pharmacological studies, proof of clinical efficacy, and safety of phytomedicines.