多西米新通过靶向FAK降低Snail和Cyclin D1的稳定性以抑制非小细胞肺癌（NSCLC）进展

Diosmin reduces the stability of Snail and Cyclin D1 by targeting FAK to inhibit NSCLC progression

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 Dec

作者: Chenkang Ma, Mengxia Dan, Ying Wang, Chenying Shu, Min Jiao, Yuna Shao, Huiling Zhang, Chang Li, Yuanyuan Zeng, Jianjie Zhu, Jian-An Huang, Jianjun Li, Zeyi Liu

DOI: 10.1016/j.phymed.2024.156135

摘要

在不同肿瘤中，焦点粘附激酶（FAK）是一种非受体酪氨酸激酶，其表达上调，因此成为癌症治疗的潜在靶点。然而，FAK激酶抑制剂的研发面临诸多挑战。因此，亟需迅速鉴定新的有效FAK激酶抑制剂。通过分子对接筛选到靶向FAK的小分子，并通过表面等离子共振（SPR）和细胞热稳定性分析（CETSA）验证。我们采用CCK-8、克隆形成、EdU和Transwell等体外实验，以及细胞周期分析，研究FAK激酶抑制剂的药理作用。采用免疫共沉淀、RNA拉下和RNA免疫沉淀等技术探讨其分子机制。结果显示，Diosmin（Dio）是一种FAK抑制剂，能在肺腺癌中展现抗增殖和抗转移作用。机制上，Dio通过抑制FAK的催化活性，阻断肿瘤细胞增殖和转移。Dio促使泛素-蛋白酶体途径降解Cyclin D1，同时通过减少Snail的mRNA稳定性，抑制上皮-间质转化（EMT）过程，从而逆转肿瘤的转移。并在小鼠异种移植模型中验证了Dio对肺腺癌的抑制效果。这些结果支持FAK在肺腺癌中通过稳定Cyclin D1和Snail促进肿瘤的作用，并表明Dio是一种具有潜力的FAK抑制候选药物。

Abstract

In different tumours, focal adhesion kinase (FAK), a nonreceptor tyrosine kinase, is upregulated and hence, it represents a promising target for cancer therapy. However, the development of FAK kinase inhibitors has faced a number of challenges. It is therefore imperative that new, effective FAK kinase inhibitors be identified promptly.Small molecules that target FAK were identified through molecular docking and validated through surface plasmon resonance (SPR) and cell thermal shift analysis. We investigated the pharmacological effects of FAK kinase inhibitors using CCK-8, colony formation, EdU, and Transwell assays and cell cycle analysis. The molecular mechanism was determined via methods such as coimmunoprecipitation, RNA pull-down and RNA immunoprecipitation.Here, we confirmed that diosmin (Dio) is an inhibitor of FAK and demonstrated its anti-proliferative and anti-metastatic effects in lung adenocarcinoma. Mechanistically, Dio inhibited tumour proliferation and metastasis by impeding the catalytic activity of FAK. Dio activated the ubiquitin proteasome pathway to induce Cyclin D1 degradation, while inhibiting tumour proliferation and reversing the epithelial mesenchymal transition (EMT) process by reducing the mRNA stability of Snail, thereby inhibiting cancer metastasis. In addition, the inhibitory effect of Dio on lung adenocarcinoma was validated in a mouse xenograft model.These results support the tumour-promoting role of FAK in lung adenocarcinoma by stabilizing Cyclin D1 and Snail and suggest that Dio is a promising candidate for FAK inhibition.