苦杏仁苷在KRAS突变非小细胞肺癌细胞生长和肿瘤免疫中的作用及机制  

Role and mechanism of amygdalin in cell growth and tumor immunity in KRAS mutated non-small cell lung cancer cells

作　　者：刘奎[1] LIU Kui(Department of Thoracic and Cardiovascular Surgery,The Fourth People’s Hospital of Zigong City,Zigong 64300,Sichuan,China)

机构地区：[1]自贡市第四人民医院胸心大血管外科,四川自贡643000

出　　处：《皖南医学院学报》2025年第2期107-111,共5页Journal of Wannan Medical College

基　　金：四川省医学会重症医学(国瑞)专项科研课题(18PJ1165)。

摘　　要：目的:探讨苦杏仁苷对Kirsten鼠肉瘤基因(KRAS)突变非小细胞肺癌(NSCLC)细胞生长和肿瘤免疫的影响,并探讨可能的潜在机制。方法:用不同浓度(10、50、100、150、200μg/mL)的苦杏仁苷处理KRAS突变的人NSCLC细胞系A54924 h。同时,用程序性死亡配体1(PD-L1)的选择性激活剂干扰素γ(IFNγ)和Yes相关蛋白(YAP)抑制剂维替洛芬(VP)联合150μg/mL苦杏仁苷处理A549细胞24 h进行挽救实验或协同实验。采用MTT法检测细胞活力;EdU incorporation assay法检测细胞增殖;流式细胞术检测细胞凋亡;伤口愈合实验和Transwell实验检测细胞迁移与侵袭;蛋白免疫印迹检测PD-L1、YAP、磷酸化YAP(p-YAP)、转录共激活因子(TAZ)蛋白的表达;实时定量聚合酶链式反应(qRT-PCR)检测CTGF和CYR61 mRNA的表达。结果:苦杏仁苷能够以剂量依赖性的方式降低A549细胞的活力(P<0.05)。50、100、150μg/mL的苦杏仁苷能够抑制A549细胞的增殖、迁移和侵袭,促进其凋亡(P<0.05)。苦杏仁苷能够抑制A549细胞中PD-L1的表达,且这一作用能够在一定程度上被IFNγ消除(P<0.05)。苦杏仁苷能够抑制YAP和TAZ蛋白表达,促进YAP磷酸化,抑制Hippo-YAP信号通路下游靶标CTGF和CYR61 mRNA的表达,且苦杏仁苷与VP联合使用能够协同下调A549细胞中PD-L1的表达(P<0.05)。结论:苦杏仁苷能够抑制KRAS突变型NSCLC细胞的增殖、迁移和侵袭,并诱导细胞凋亡,这可能是通过调节Hippo-YAP/PD-L1轴以调节肿瘤免疫抑制来实现。Objective:To investigate the effects of amygdalin on the growth and tumor immunity of Kirsten rat sarcoma gene(KRAS)mutated non-small cell lung cancer(NSCLC)cells and its possible underlying mechanisms.Methods:Different concentrations of amygdalin(10,50,100,150,and 200μg/mL)were used to treat the human KRAS-mutated NSCLC cell line A549 for 24 hours.Rescue or synergy experiments were performed by treating the cell line with a combination of interferon gamma(IFNγ),a selective activator of programmed death ligand 1(PD-L1),and Verteporfin(VP),a Yes-associated protein(YAP)inhibitor,along with 150μg/mL amygdalin for 24 hours.MTT assay was used to measure cell viability.Edu incorporation assay was employed to assess cell proliferation.Flow cytometry was applied to detect cell apoptosis;Wound-healing assay and Transwell assay were respectively conducted to evaluate cell migration and invasion.Western blotting was performed to analyze the protein expression of PD-L1,YAP,phosphorylated YAP(p-YAP),and transcriptional co-activator with PDZ-binding motif(TAZ).qRT-PCR was utilized to detect the mRNA expression of CTGF and CYR61.Results:Amygdalin reduced the viability of A549 cells in a dose-dependent manner(P<0.05).Amygdalin(at 50,100,and 150μg/mL)inhibited the proliferation,migration and invasion of A549 cells and promoted their apoptosis(P<0.05).Amygdalin suppressed the expression of PD-L1 in A549 cells,and this effect could be partially abolished by IFNγ(P<0.05).Amygdalin inhibited the expression of YAP and TAZ proteins,promoted the phosphorylation of YAP,and suppressed the mRNA expression of Hippo-YAP signaling downstream targets CTGF and CYR61.Combined use of amygdalin with VP synergistically downregulated the expression of PD-L1 in A549 cells(P<0.05).Conclusion:Amygdalin can inhibit the proliferation,migration,and invasion of KRAS-mutated NSCLC cells as well as induce the cellular apoptosis,which is potentially associated with regulation of the Hippo-YAP/PD-L1 axis to modulate tumor immune suppression.

关 键 词：苦杏仁苷 KRAS突变非小细胞肺癌细胞 程序性死亡配体1 Yes相关蛋白/转录共激活因子 

分 类 号：R285[医药卫生—中药学] R734.2[医药卫生—中医学]