Feiyiliu Mixture sensitizes EGFRDel19/T790M/C797S mutant non-small cell lung cancer to osimertinib by attenuating the PRC1/Wnt/EGFR pathway

Abstract

Introduction: Osimertinib is a potent epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) for the treatment of patients with EGFR-mutant non-small cell lung cancer (NSCLC). However, the emergence of acquired resistance due to the EGFR-Del19/T790M/C797S mutation limits the clinical application of osimertinib. Feiyiliu Mixture (FYLM), a clinical experience formula of Chinese medicine, was used to treat lung cancer with good clinical efficacy. In this study, we aimed to investigate the mechanism by which Feiyiliu Mixture delays osimertinib resistance in EGFR-mutant cell lines and EGFR-mutant cell tumor-bearing mice.Methods: The osimertinib-resistant cell models were established in mouse Lewis lung carcinoma (LLC) cells transfected with EGFR-Del19/T790M/C797S mutant lentivirus. In cell experiments, after 48 h of treatment with Feiyiliu Mixture-containing serum, MTT assay was used to detect the relative cell viability, and western blotting was used to detect EGFR protein phosphorylation expression. In animal experiments, C57BL/6J mice were subcutaneously injected with Lewis lung carcinoma cells stably expressing EGFR-Del19/T790M/C797S mutations to construct a xenograft model. After 2 weeks of Feiyiliu Mixture and/or osimertinib treatment, the expression of proliferation-related, apoptosis-related and PRC1/Wnt/EGFR pathway markers was detected by real-time qPCR, western blotting and immunohistochemistry.Results: The results showed that when combined with osimertinib, Feiyiliu Mixture synergistically reduces proliferation and increases apoptosis to improve drug resistance. In vitro, Feiyiliu Mixture-containing serum reduced the EGFR phosphorylation. In vivo, Feiyiliu Mixture downregulated the expression of cyclin B1 and Bcl-2 while upregulating the level of cleaved Caspase-3 protein, indicating that Feiyiliu Mixture promotes apoptosis. Furthermore, Feiyiliu Mixture reduced the expression of p-EGFR, p-Akt, PRC1 and Wnt pathway-related proteins such as β-catenin, c-Myc and c-Jun.Conclusion: The present study identified that Feiyiliu Mixture inhibited PRC1/Wnt/EGFR pathway activation, reduced proliferation, and promoted apoptosis, thereby increasing the sensitivity of EGFR-mutant non-small cell lung cancer to osimertinib. Our study provided a new idea for Chinese medicine to play a role in enhancing efficacy and reducing toxicity in the treatment of non-small cell lung cancer.

The clinicopathological features and possible physiological mechanisms of only EGFR-T790M primary mutation in lung adenocarcinoma patients

“…This is consistent with Reguart et al 32 and Ye et al 33 who reported that after 1-2 years, patients will inevitably develop resistance to EGFR-TKIs, resulting in secondary T790M mutation. With the increase of EGFR drug resistance, for example, the third generation TKI osimertinib is selective to activation mutation and T790M drug resistance, and the patient may have a mutation of C797S after 9-13 months' of treatment, and at present, there is no treatment for the drug resistance mediated by the triple mutation of 19Del/T790M/C797S, so it is necessary to further study the drug resistance mechanism of EGFR mutation and nd new targeted drug therapy [34][35][36][37] ; Therefor, more TKIs are approved to enter the clinic by FAD, which increases the economic pressure and drug toxicity of patients, such as zhu et al whose results showed that serious adverse events (SAE) induced by two different strategies of targeted therapy is 60% and 28% respectively 16 . Certainly, the solution to TKIs drug resistance is gradually being overcome, and EGFR is targeted through allosteric mechanism to overcome the drug resistance in ATP binding sites 38 ; A new class of EGFR PROTACs degrading agent based on pomalidomide was developed by O. Aboelez et al, which has good activity on both wild-type and mutant EGFR, and its anti-tumor cell proliferation effect has been con rmed in lung cancer cells in vitro 39 .…”

Association of lung-intestinal microecology and lung cancer therapy

“…YYD inhibited the proliferation of NSCLC cells by inhibiting the EGFR-PI3K-AKT signaling pathway [ 108 ]. Feiyiliu Mixture could reduce the phosphorylation of EGFR and down-regulate the expression of cyclin, and up-regulate the level of cleaved Caspase-3 protein to promote cell apoptosis [ 109 ]. Therefore, lung cancer treatment should consider modulating intestinal function as well.…”

Dysregulated Signalling Pathways Driving Anticancer Drug Resistance

“…Moreover, osimertinib is a third-generation powerful EGFR-TKI used to treat NSCLC patients with EGFR mutations. The therapeutic use of osimertinib is nonetheless restricted by the emergence of acquired resistance associated with the triple mutation Del19/T790M/C797S in EGFR [63]. Furthermore, mutations at either V550 (a gatekeeper residue) or C552 (hinge-1 residue) in the kinase domain of fibroblast growth factor receptor 4 (FGFR4) prevent fisogatinib (a potent and selective FGFR4 inhibitor) from interacting with the FGFR4's ATP binding site, resulting in acquired clinical resistance to fisogatinib in patients with HCC [64].…”