In vitro study of CAR-NK cell immunotherapy targeting c-Met for gastric cancer
10.3969/j.issn.1673-9701.2025.20.008
- VernacularTitle:靶向c-Met的CAR-NK细胞免疫治疗胃癌的体外研究
- Author:
Huifei LU
1
;
Liqi MAO
1
;
Guijun WEI
1
Author Information
1. 湖州市第一人民医院消化内科,浙江湖州 313000
- Publication Type:Journal Article
- Keywords:
Cellular-mesenchymal epithelial transition factor;
Chimeric antigen receptor natural killer cells;
Cellular immunotherapy;
Gastric cancer
- From:
China Modern Doctor
2025;63(20):31-34,56
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigates the inhibitory and cytotoxic effects of chimeric antigen receptor-engineered natural killer(CAR-NK)cells targeting cellular-mesenchymal epithelial transition factor(c-Met)against gastric cancer(GC)cells in vitro.Methods c-Met expression at protein and mRNA levels was evaluated in GC cell lines(MKN-45,GTL-16,NCI-N87)and gastric mucosal epithelial cells(GES-1)by using flow cytometry(FCM),Western blot,and reverse transcription quantitative polymerase chain reaction.Immunofluorescence assays were conducted to confirm c-Met CAR-mediated specificity toward c-Met-overexpressing GTL-16 cells.Lentiviral vectors were utilized to construct c-Met-targeting CAR-NK cells,with transduction efficiency and phenotypic integrity verified by FCM.Cytotoxic activity against GC cells and normal epithelial cells was quantified via lactate dehydrogenase(LDH)release assays and cytokine secretion measured by enzyme-linked immunosorbent assay.Results c-Met exhibited significantly higher expression in GTL-16 and MKN-45 cells at both transcriptional and protein levels compared to NCI-N87 and GES-1 cells(P＜0.05).CAR-NK cells demonstrated a transduction efficiency of(14.97±0.25)％ without altering NK-92 cell phenotypes.Immunofluorescence confirmed specific binding of c-Met CAR-expressing Lenti-X-293T cells to GTL-16 targets.LDH assays revealed enhanced cytotoxicity of c-Met CAR-NK cells against c-Met-high GC cells(GTL-16,MKN-45)versus untransduced NK-92 controls(P＜0.05).At a 10∶1 effector-to-target ratio,c-Met CAR-NK cells co-cultured with c-Met-high GC cells secreted significantly elevated tumor necrosis factor(TNF-α)and interferon(IFN)-γ(P＜0.05).Conclusion The c-Met CAR-NK cells constructed in this study have the ability to specifically recognize and target to kill gastric cancer cells with high expression of c-Met,and have a strong killing effect function,which can release more TNF-α and IFN-γ.
