葫芦素B抑制奥希替尼耐药的非小细胞肺癌细胞迁移侵袭的机制研究  

Cucurbitacin B regulates the resistance of osimertinib by inhibiting cell migration and invasion in non-small cell lung cancer

作　　者：王婷[1] 钱强 陈荣 何子珩 曹鹏[2,3] 蔡雪婷[1] 崔鹤 WANG Ting;QIAN Qiang;CHEN Rong;HE Zi-heng;CAO Peng;CAI Xue-ting;CUI He(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing 210028,China;School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;State Key Laboratory on Technologies for Chinese Medicine Pharmaceutical Process Control and Intelligent Manufacture,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学附属中西医结合医院,江苏南京210028 [2]南京中医药大学药学院,江苏南京210023 [3]南京中医药大学,中药制药过程控制与智能制造技术全国重点实验室,江苏南京210023

出　　处：《药学学报》2025年第3期731-738,共8页Acta Pharmaceutica Sinica

基　　金：国家重点研发计划(2023YFC2308200);国家自然科学基金资助项目(82125037,82273153);江苏省临床中药学交叉医学创新中心(CXZX202225)。

摘　　要：非小细胞肺癌(non-small cell lung cancer,NSCLC)是肺癌的主要病理类型,奥希替尼作为第三代表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitor,EGFR-TKI)靶向药物,能有效延长EGFR突变NSCLC患者的无进展生存期。然而,耐药性限制了奥希替尼的疗效,而中医药能有效延缓EGFR-TKIs的耐药性。本研究以葫芦素B(cucurbitacin B,CuB)为研究对象,通过细胞活力检测和细胞迁移、侵袭实验分析葫芦素B在奥希替尼耐药NSCLC细胞中的药效作用;使用公共数据库筛选葫芦素B在奥希替尼耐药NSCLC细胞中的潜在靶点,并通过分子对接、细胞热位移实验(cellular thermal shift assay,CETSA)和微量热泳动实验(microscale thermophoresis,MST)验证葫芦素B与潜在靶点之间的相互作用;免疫印迹实验检测葫芦素B对靶点下游信号通路的影响。结果显示,葫芦素B能显著降低奥希替尼耐药NSCLC细胞的生长活力,抑制细胞的迁移和侵袭能力;机制上,葫芦素B通过结合AXL进而抑制其下游细胞外信号调节激酶(extracellular regulated protein kinases,ERK)和蛋白激酶B(protein kinase B,AKT)的激活。综上所述,葫芦素B具有缓解NSCLC奥希替尼治疗抵抗的潜在作用,其可通过下调AXL-ERK/AKT轴的异常激活,抑制耐药细胞的迁移、侵袭。本研究为葫芦素B治疗NSCLC奥希替尼靶向耐药提供了临床前体外药效的依据,也为临床联合用药开发提供了理论支持。Non-small cell lung cancer(NSCLC)is the primary pathological type of lung cancer.Osimertinib,as a third-generation EGFR-TKI(epidermal growth factor receptor-tyrosine kinase inhibitor)targeted drug,effectively prolong the progression-free survival of patients with EGFR-mutated NSCLC.However,drug resistance limits the efficacy of osimertinib.Traditional Chinese medicine can effectively delay the resistance to EGFR-TKIs.In this study,Cucurbitacin B(CuB)was investigated to analyze its pharmacological effects in osimertinib-resistant NSCLC cells through cell viability,migration,and invasion experiments.Public databases were used to screen potential targets of CuB in osimertinib-resistant NSCLC cells,and the interactions between CuB and potential targets were verified through molecular docking,cellular thermal shift assay(CETSA),and microscale thermophoresis(MST).Western blot was used to detect the effects of CuB on downstream pathways of the targets.The results showed that CuB significantly reduced the proliferation activity of osimertinib-resistant NSCLC cells and inhibited the migration and invasion abilities of tumor cells.Mechanistically,CuB inhibited the expression of ERK and AKT molecules by binding to the tyrosine kinase receptor AXL.In summary,CuB exhibits resistance to osimertinib-resistant NSCLC by inhibiting the migration and invasion of resistant cells through regulating the abnormal activation of the AXL-ERK/AKT axis.This study provides a basis for the pre-clinical in vitro efficacy of CuB in the treatment of osimertinib targeted resistance in NSCLC and theoretical support for the development of clinical drug combinations.

关 键 词：非小细胞肺癌 耐药 奥希替尼 葫芦素B AXL 

分 类 号：R966[医药卫生—药理学]