Vitexicarpin通过影响IMPDH2靶向c-Myc泛素化抑制结直肠癌的恶性进展

Vitexicarpin suppresses malignant progression of colorectal cancer through affecting c-Myc ubiquitination by targeting IMPDH2

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PHYTOMEDICINE

影响因子:8.3

分区:医学1区 Top / 药物化学1区全科医学与补充医学1区药学1区植物科学1区

发表日期:2024 Sep

作者: Xiao-Jing Ding, Xue-Mei Cai, Qian-Qian Wang, Ning Liu, Wei-Long Zhong, Xiao-Nan Xi, Ya-Xin Lu

DOI: 10.1016/j.phymed.2024.155833

摘要

结直肠癌（CRC）是继发性癌症相关死亡的第二大原因，具有广泛的侵袭和转移潜能。此前的研究显示，从Vitex rotundifolia果实提取的vitexicarpin可以阻碍肿瘤进展。然而，涉及CRC治疗的分子机制尚未完全明确。本研究旨在探讨vitexicarpin的抗癌活性、靶点及其在CRC中的分子机制，希望为CRC患者提供新的治疗策略。我们通过MTT、克隆形成、EDU、细胞周期和凋亡检测，以及肿瘤异种移植模型，评估了vitexicarpin对CRC的影响。采用CETSA、无标记定量蛋白质组学和Biacore技术识别其靶点。利用WB、Co-IP、泛素化检测、免疫荧光、分子对接、MST和细胞转染，研究了vitexicarpin在CRC细胞中的作用机制。此外，通过分析TCGA和GEPIA数据库及临床样本中靶蛋白的表达模式和相关性，进一步验证了其机制。最后，利用伤口愈合、Transwell、尾静脉注射模型和组织切片染色，展示了vitexicarpin在体内外的抗转移作用。结果显示，vitexicarpin通过直接结合次黄嘌呤单磷酸脱氢酶2（IMPDH2）发挥抗癌作用，并通过破坏IMPDH2与c-Myc的相互作用，促进c-Myc的泛素化，从而抑制上皮-间充质转化（EMT）。vitexicarpin阻碍CRC细胞迁移和侵袭，逆转EMT过程，作用在体外和体内均得到验证。此外，通过IMPDH2的过表达和敲低，验证了这些作用机制。这些结果表明，vitexicarpin通过调控IMPDH2与c-Myc的相互作用，抑制CRC的增殖和转移，为CRC的治疗提供了潜在的分子靶点，并揭示了c-Myc在肿瘤中的新调控机制。

Abstract

Colorectal cancer (CRC) is the second most common cause of cancer-related mortality and is characterised by extensive invasive and metastatic potential. Previous studies have shown that vitexicarpin extracted from the fruits of Vitex rotundifolia can impede tumour progression. However, the molecular mechanisms involved in CRC treatment are still not fully established.Our study aimed to investigate the anticancer activity, targets, and molecular mechanisms of vitexicarpin in CRC hoping to provide novel therapies for patients with CRC.The impact of vitexicarpin on CRC was assessed through various experiments including MTT, clone formation, EDU, cell cycle, and apoptosis assays, as well as a tumour xenograft model. CETSA, label-free quantitative proteomics, and Biacore were used to identify the vitexicarpin targets. WB, Co-IP, Ubiquitination assay, IF, molecular docking, MST, and cell transfection were used to investigate the mechanism of action of vitexicarpin in CRC cells. Furthermore, we analysed the expression patterns and correlation of target proteins in TCGA and GEPIA datasets and clinical samples. Finally, wound healing, Transwell, tail vein injection model, and tissue section staining were used to demonstrate the antimetastatic effect of vitexicarpin on CRC in vitro and in vivo.Our findings demonstrated that vitexicarpin exhibits anticancer activity by directly binding to inosine monophosphate dehydrogenase 2 (IMPDH2) and that it promotes c-Myc ubiquitination by disrupting the interaction between IMPDH2 and c-Myc, leading to epithelial-mesenchymal transition (EMT) inhibition. Vitexicarpin hinders the migration and invasion of CRC cells by reversing EMT both in vitro and in vivo. Additionally, these results were validated by the overexpression and knockdown of IMPDH2 in CRC cells.These results demonstrated that vitexicarpin regulates the interaction between IMPDH2 and c-Myc to inhibit CRC proliferation and metastasis both in vitro and in vivo. These discoveries introduce potential molecular targets for CRC treatment and shed light on new mechanisms for c-Myc regulation in tumours.