中药新药与临床药理2026，Vol.37Issue(1)：12-21,10.DOI:10.19378/j.issn.1003-9783.2026.01.002
复方蜥蜴散干预肿瘤细胞干性对胃癌顺铂耐药的增敏提效作用机制
Mechanism of Compound Xiyi San in Sensitizing Gastric Cancer to Cisplatin by Trageting Tumor Stemness
摘要
Abstract
Objective To investigate the mechanism by which Compound Xiyi San(CXS)enhances cisplatin sensitivity in gastric cancer by interfering with tumor stemness via the interferon-γ(IFN-γ)/nuclear factor-κB(NF-κB)/SNAIL axis.Methods(1)MKN45 cells and MKN45/DDP cisplatin-resistant cells in the logarithmic growth phase were treated with different concentrations(0,0.2,0.4,0.8,1.6,3.2 μg·mL-1)of cisplatin(DDP)for 48 hours.Cell proliferation capacity and the resistance index were measured using the CCK-8 assay.The cancer stem cell characteristics of MKN45 and MKN45/DDP cells were validated using the serum-free suspension sphere formation assay.(2)MKN45/DDP cells in the logarithmic growth phase were subcutaneously inoculated into the right anterior axilla of male BALB/c nude mice to establish a cisplatin-resistant gastric cancer xenograft model.Successfully modeled mice were randomly divided into four groups(n=8 per group):model group,cisplatin group,CXS group,and combination group.The model group received an equal volume of normal saline by gavage;the cisplatin group received intraperitoneal injections of 0.002 g·kg-1 cisplatin twice weekly;the CXS group received 2.8 g·kg-1 CXS aqueous extract by gavage twice daily;the combination group received both CXS aqueous extract(2.8 g·kg-1,twice daily)and cisplatin(0.002 g·kg-1,twice weekly).All interventions continued for 4 weeks.Tumor mass was weighed,and tumor volume was calculated.Pathological changes in tumor tissue were observed by HE staining;apoptosis was detected by TUNEL assay;protein expression of Ki67,EpCAM,and CD44 in tumor tissue was determined by immunohistochemistry;and protein expression levels of multidrug resistance-associated protein 1(MRP1),P-glycoprotein(P-gp),EpCAM,CD44,IFN-γ,p65,phosphorylated p65(p-p65),and SNAIL were measured by Western Blot.Results(1)Compared with MKN45 cells,the survival rates of MKN45/DDP cells in the 0.8,1.6,and 3.2 μg·mL-1 cisplatin groups were significantly increased(P＜0.05,P＜0.01).The IC50 for MKN45 cells was 0.96 μg·mL-1,and for MKN45/DDP cells it was 2.26 μg·mL-1,resulting in a resistance index of 2.36 for MKN45/DDP cells.Compared with MKN45 cells,MKN45/DDP cells exhibited a significantly enhanced sphere-forming ability(P＜0.01).(2)In the model group,tumor cells showed irregular morphology,tight arrangement,large nuclei,and pathological mitotic figures.Compared with the model group,all treatment groups showed significantly reduced tumor volume(P＜0.01)and tumor mass(P＜0.01)in MKN45/DDP xenograft nude mice;mitotic figures were reduced,cell density decreased,arrangement became looser,and necrotic cells were observed;tumor cell apoptosis rates were significantly increased(P＜0.01);Ki67 positive expression levels in tumor tissue were significantly decreased(P＜0.05,P＜0.01);protein expression levels of stemness markers EpCAM and CD44,and drug resistance-related proteins MRP1 and P-gp were significantly decreased(P＜0.05,P＜0.01);and protein expression levels of p-p65/p65,IFN-γ,and SNAIL in tumor tissue were significantly decreased(P＜0.01).Compared with the cisplatin group,the tumor volume and tumor mass of MKN45/DDP xenograft nude mice in the CXS group and the combination group were significantly reduced(P＜0.01);significantly increased apoptosis rates(P＜0.01);significantly decreased Ki67 positive expression(P＜0.05,P＜0.01),and significantly lower protein expression levels of CD44,MRP1,P-gp,IFN-γ,and SNAIL(P＜0.05,P＜0.01);the combination group also showed significantly decreased EpCAM and p-p65/p65 protein expression levels(P＜0.01).Conclusion Compound Xiyi San effectively inhibits tumor growth,suppresses cell proliferation,and promotes apoptosis in nude mice bearing cisplatin-resistant MKN45/DDP gastric cancer xenografts.This effect may be related to the suppression of tumor stemness and subsequent reduction of cisplatin resistance through regulation of the IFN-γ/NF-κB/SNAIL axis.关键词
复方蜥蜴散/胃癌/顺铂耐药/肿瘤细胞干性/IFN-γ/NF-κB/SNAIL轴/MKN45/DDP细胞/裸鼠Key words
Compound Xiyi San/gastric cancer/cisplatin resistance/tumor stemness/IFN-γ/NF-κB/SNAIL axis/MKN45/DDP cells/nude mice分类
医药卫生引用本文复制引用
刘彩月,张泽洋,李铮,李卫强..复方蜥蜴散干预肿瘤细胞干性对胃癌顺铂耐药的增敏提效作用机制[J].中药新药与临床药理,2026,37(1):12-21,10.基金项目
国家自然科学基金项目(82260916) （82260916）
宁夏医科大学科学研究资助项目(XZ2023012). （XZ2023012）
