常春藤皂苷元调控IL-6抑制肝癌细胞增殖的免疫机制  

Immune mechanism of inhibition of hepatocellular carcinoma cells by Hederagenin via regulating IL-6

作　　者：崔文超 刘明远[1] 关宝生[2] 曾佳 田亚妮 白雪(指导)[3] CUI Wenchao;LIU Mingyuan;GUAN Baosheng;ZENG Jia;TIAN Yani;BAI Xue(Jiamusi University of Basic Medical College,Jiamusi 154007,China)

机构地区：[1]佳木斯大学基础医学院,佳木斯154007 [2]佳木斯大学公共卫生学院,佳木斯154007 [3]哈尔滨医科大学附属第二医院,哈尔滨150001

出　　处：《中国免疫学杂志》2023年第9期1864-1871,共8页Chinese Journal of Immunology

基　　金：黑龙江省自然科学基金项目(YQ2020H001);黑龙江省教育科学“十三五”规划课题(GBD1317135)项目。

摘　　要：目的:揭示常春藤皂苷元(HD)对IL-6的调控作用及其抑制肝癌细胞增殖的免疫机制。方法:(1)生物信息学分析受IL-6调控的免疫途径:利用GEO_(2)R工具分析基因芯片数据集GSE14632中的差异表达基因,Immport数据库下载免疫基因,筛选二者交集基因,KOBAS数据库对交集基因进行京都基因与基因组百科全书(KEGG)通路富集分析。(2)实验验证HD对IL-6的调控作用:Western blot检测HD(0、30、60、120μg/ml)干预下HepG2细胞内IL-6蛋白的相对表达量;构建H22荷瘤小鼠模型,荷瘤小鼠被随机分为5个实验组,每组12只:对照组、阳性环磷酰胺(CTX)25 mg/kg组、常春藤皂苷元低剂量组(100 mg/kg)、中剂量组(200 mg/kg)和高剂量组(400 mg/kg),小鼠腹腔注射给药,连续给药14 d,第15天所有动物称重后处死并取肿瘤组织,Western blot检测各实验组肿瘤组织中IL-6、KRAS、RAF1、MEK2、p-ERK1/2、p-JNK、AKT、NF-κB p50、PD-L1和p-STAT3蛋白相对表达。结果:生物信息学分析发现MAPK信号通路、PD-L1的表达和PD-1检查点通路受到IL-6信号途径的调控;体外实验结果表明,与对照组相比,IL-6在HD(30、60、120μg/ml)干预组蛋白相对表达呈现下降趋势(P<0.01);体内实验结果表明,与对照组相比,IL-6、KRAS、RAF1、MEK2、p-ERK1/2、p-JNK、AKT、NF-κB p50、PD-L1和p-STAT3在HD(100、200、400 mg/kg)干预组蛋白相对表达总体呈现下降趋势(P<0.01)。结论:HD可能通过抑制IL-6蛋白表达,进而调控肝癌细胞内PD-1/PD-L1和MAPK免疫信号通路活性,导致肝癌的适应性免疫反应受到损伤,从而抑制癌细胞的增殖。Objective:To reveal the regulatory role of Hederagenin on IL-6 and its possible immune mechanism for inhibiting the proliferation of hepatocellular carcinoma cells.Methods:①Bioinformatics was used to analyze the immune pathways regulated by IL-6:the GEO2R tool was used to analyse differentially expressed genes in GeneChip dataset GSE14632 and the Immport database was used to download immune genes,intersecting genes were screened between differentially expressed genes and immune genes.The KOBAS database was used to perform KEGG enrichment analysis of intersecting genes.②Experimental validation of the regulation of IL-6 by HD:Western blot was used to detect the relative expression of IL-6 protein in HepG2 cells under HD(0,30,60 and 120μg/ml)intervention.H22 tumour-bearing mice were randomly divided into 5 groups,12 cases in each group:control group;CTX group,25 mg/kg;HD low dose group,100 mg/kg;HD medium dose group,200 mg/kg;HD high dose group,400 mg/kg.Mice were administered intraperitoneally for 14 days.On day 15,all animals were weighed and executed and the tumours were removed.Western blot was used to detect the relative expression of IL-6,KRAS,RAF1,MEK2,p-ERK1/2,p-JNK,AKT,NF-κB p50,PD-L1 and p-STAT3 proteins in the tumours of each group.Results:Bioinformatics analysis revealed that MAPK signaling pathway,PD-L1 expression and PD-1 checkpoint pathway were regulated by IL-6;the result of in vitro experiment showed that the relative protein expression of IL-6 in HD(30,60 and 120μg/ml)intervention groups showed a decreasing trend compared to control group(P<0.01);the results of in vivo experiment showed that the relative proteins expression of IL-6,KRAS,RAF1,MEK2,p-ERK1/2,p-JNK,AKT,NF-κB p50,PD-L1 and p-STAT3 in HD(100,200 and 400 mg/kg)intervention groups showed an overall decreasing trend compared to control group(P<0.01).Conclusion:HD may regulate the activity of PD-1/PD-L1 and MAPK immune signaling pathways in liver cancer cells by inhibiting the expression of IL-6 protein,resulting in the damage of ada

关 键 词：肝细胞癌 常春藤皂苷元 生物信息学 IL-6 MAPK信号通路 PD-1/PD-L1 

分 类 号：R285[医药卫生—中药学]