柚皮素通过阻断 IL18/IL18RAP 信号轴来阻碍胃癌细胞与癌相关成纤维细胞之间的串扰。

Eupafolin hinders cross-talk between gastric cancer cells and cancer-associated fibroblasts by abrogating the IL18/IL18RAP signaling axis.

机构信息

Henan Provincial Clinical Research Center for Pediatric Diseases, Henan Key Laboratory of Pediatric Genetics and Metabolic Diseases, Children's Hospital Affiliated to Zhengzhou University, Henan Children's Hospital, Zhengzhou Children's Hospital, Zhengzhou, China; Department of Cardiothoracic Surgery, Children's Hospital Affiliated to Zhengzhou University, Henan Children's Hospital, Zhengzhou Children's Hospital, Zhengzhou, China.

Henan Provincial Clinical Research Center for Pediatric Diseases, Henan Key Laboratory of Pediatric Genetics and Metabolic Diseases, Children's Hospital Affiliated to Zhengzhou University, Henan Children's Hospital, Zhengzhou Children's Hospital, Zhengzhou, China; Department of Pathology, Children's Hospital Affiliated to Zhengzhou University, Henan Children's Hospital, Zhengzhou Children's Hospital, Zhengzhou, China.

出版信息

Phytomedicine. 2024 Nov;134:155984. doi: 10.1016/j.phymed.2024.155984. Epub 2024 Aug 31.

DOI:10.1016/j.phymed.2024.155984

PMID:39265444

Abstract

BACKGROUND

Cancer-associated fibroblasts (CAFs) are involved in the progression of gastric cancer (GC) as a critical component of the tumor microenvironment (TME), yet specific interventions remain limited. Natural products hold a promising application prospect in the field of anti-tumor in view of their high activity and ease of binding with biological macromolecules. However, the role of natural products in modulating the cross-talk between CAFs and GC cells has not been fully investigated.

PURPOSE

The aim of this study was to identify a potential therapeutic target in CAFs and then screen for natural small molecule drugs with anti-tumor activity against this target.

METHODS

Integrating bioinformatics analysis of public databases and experimental validation of human samples and cell lines to identify a candidate target in CAFs. Molecular docking and biolayer interferometry technique were utilized for screening potential natural small molecule drugs. The efficacy and underlying mechanisms of the candidates were explored in vitro and in vivo through techniques such as lentiviral infection, cell spheroids culture, immunoprecipitation and cells-derived xenografts.

RESULTS

IL18 receptor accessory protein (IL18RAP) was found to be overexpressed in CAFs derived from GC tissues and facilitated the protumor function of CAFs on GC. Based on virtual screening and experimental validation, we identified a natural product, eupafolin, that interfered with IL18 signaling. Phenotyping studies confirmed that the proliferation, spheroids formation and tumorigenesis of GC cells facilitated by CAFs were greatly attenuated by eupafolin both in vitro and in vivo. Mechanistically, eupafolin impeded the formation of IL18 receptor (IL18R) complex by directly binding to IL18RAP, thus blocking IL18-mediated nuclear factor kappa B (NF-κB) activation and reduced the synthesis and secretion of IL6 in CAFs. As a consequence, it inactivated signal transducer and activator of transcription 3 (STAT3) in GC cells.

CONCLUSION

This study provides new evidence that IL18 signaling regulates the cross-talk between GC cells and CAFs. And it highlights a novel pharmacological role of eupafolin in inhibiting IL18 signaling, thereby curbing the development of GC via modulating CAFs.

摘要

背景

癌症相关成纤维细胞（CAFs）是肿瘤微环境（TME）的重要组成部分，参与胃癌（GC）的进展，但特定干预措施仍然有限。鉴于天然产物具有高活性和易于与生物大分子结合的特点，在抗肿瘤领域具有广阔的应用前景。然而，天然产物在调节 CAFs 与 GC 细胞之间的串扰方面的作用尚未得到充分研究。

目的

本研究旨在确定 CAFs 中的一个潜在治疗靶点，然后筛选针对该靶点具有抗肿瘤活性的天然小分子药物。

方法

整合公共数据库的生物信息学分析和人样本及细胞系的实验验证，以确定 CAFs 中的候选靶点。利用分子对接和生物层干涉技术筛选潜在的天然小分子药物。通过慢病毒感染、细胞球培养、免疫沉淀和细胞源性异种移植等技术，在体外和体内研究候选物的疗效和潜在机制。

结果

发现白细胞介素 18 受体辅助蛋白（IL18RAP）在源自 GC 组织的 CAFs 中过度表达，并促进 CAFs 对 GC 的促肿瘤功能。基于虚拟筛选和实验验证，我们鉴定出一种天然产物，即 eupafolin，它干扰了 IL18 信号。表型研究证实，eupafolin 在体外和体内均显著减弱 CAFs 促进 GC 细胞增殖、球状体形成和致瘤作用。在机制上，eupafolin 通过直接与 IL18RAP 结合，阻碍 IL18 受体（IL18R）复合物的形成，从而阻断 IL18 介导的核因子 kappa B（NF-κB）激活，并减少 CAFs 中 IL6 的合成和分泌。结果，它使 GC 细胞中的信号转导和转录激活因子 3（STAT3）失活。