摘要
目的:探究槲皮素(QUE)调节程序性死亡蛋白-1/程序性死亡配体-1(PD-1/PD-L1)信号通路对胃癌细胞免疫逃逸的影响。方法:使用不同浓度的QUE(0~160μmol//L)作用于胃癌细胞HGC-27,检测细胞存活率,筛选最佳药物浓度;将HGC-27细胞随机分为HGC-27组、QUE低浓度(QUE-L)组、QUE中浓度(QUE-M)组、QUE高浓度(QUE-H)组、QUE-H+pcDNANC组(过表达PD-1阴性对照)和QUE-H+pcDNA-PD-1组(过表达PD-1);CCK-8法、克隆形成实验分析细胞增殖活性;采用流式细胞术检测各组细胞凋亡率;Western blot检测PD-1/PD-L1信号通路相关蛋白表达水平;各组均与NK细胞进行共培养,构建HGC-27共培养组、QUE-L共培养组、QUE-M共培养组、QUE-H共培养组、QUE-H+pcDNA-NC共培养组和QUE-H+pcDNA-PD-1共培养组,检测NK细胞杀伤率,采用ELISA测定TNF-α、IL-2和IFN-γ表达水平。结果:相比HGC-27组,QUE-L组、QUE-M组和QUE-H组细胞存活率、克隆形成数量以及细胞中PD-1和PD-L1表达水平均逐渐降低(P<0.05),而细胞凋亡率逐渐升高(P<0.05),并且QUE-L共培养组、QUE-M共培养组和QUE-H共培养组NK细胞杀伤率及相细胞培养基中IL-2、IFN-γ和TNF-α表达水平较HGC-27共培养组也逐渐升高(P<0.05);在高浓度QUE处理HGC-27细胞的基础上过表达PD-1则逆转了以上指标的变化趋势(P<0.05)。结论:QUE可能通过下调PD-1/PD-L1信号通路抑制胃癌细胞免疫逃逸。
Objective:To investigate the effect of quercetin(QUE)on immune escape of gastric cancer cells by regulating the programmed death protein-1/programmed death ligand-1(PD-1/PD-L1)signaling pathway.Methods:Different concentrations of QUE(0~160μmol/L)were applied to gastric cancer cell HGC-27,the cell survival rate was measured to screen for the optimal drug con-centration.HGC-27 cells were stochastically grouped into HGC-27 group,QUE low concentration(QUE-L)group,QUE medium con-centration(QUE-M)group,QUE high concentration(QUE-H)group,QUE-H+pcDNA-NC group(negative control for overexpres-sion of PD-1),and QUE-H+pcDNA-PD-1 group(overexpression of PD-1).Cell proliferation activity was analyzed by CCK-8 assay and colony formation assay.Apoptosis rate was detected by flow cytometry.The expression levels of proteins related to the PD-1/PD-L1 signaling pathway were detected by Western blot.Each group was co-cultured with NK cells,and the HGC-27 co-culture group,QUE-L co-culture group,QUE-M co-culture group,QUE-H co-culture group,QUE-H+pcDNA-NC co-culture group,and QUE-H+pcDNAPD-1 co-culture group were constructed.NK cell killing rate was detected.ELISA was applied to detect the expression levels of IL-2,IFN-γand TNF-αin various cell culture media.Results:Compared with the HGC-27 group,the cell survival rate,clone formation quantity,and PD-1 and PD-L1 expression levels in the QUE-L group,QUE-M group and QUE-H group were gradually decreased(P<0.05),while the cell apoptosis rate gradually increased(P<0.05).Moreover,the NK cell killing rate and the expression levels of IL-2,IFN-γand TNF-αin the cell culture medium of the QUE-L co-culture group,QUE-M co-culture group,and QUE-H co-culture group gradually increased compared to the HGC-27 co-culture group(P<0.05).On the basis of high concentration QUE treatment of HGC-27 cells,overexpression of PD-1 reversed the trend of changes in the above indicators(P<0.05).Conclusion:QUE may inhibit immune escape of gastric cancer cells by down-regulating PD-1/PD-L1 signaling pathway.
作者
胡永进
李凤娥
HU Yongjin;LI Feng'e(Department of Oncology,Beichen Hospital Affiliated to Nankai University,Tianjin 300400,China)
出处
《中国免疫学杂志》
北大核心
2025年第8期1965-1969,共5页
Chinese Journal of Immunology
