摘要
目的 考察黄芪多糖对乳腺癌细胞的抑制作用并预测参与的通路,对关键通路进行验证,并对涉及的免疫通路进行富集分析。方法 采用CCK-8法检测不同浓度黄芪多糖(0、0.5、1.0、2.0、4.0、8.0 mg/mL)对乳腺癌MDA-MB-231细胞增殖的抑制作用;对黄芪多糖给药组和对照组进行基因表达谱测定,筛选差异表达基因,应用R语言clusterProfiler包将筛选出的差异表达基因进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析。采用Western blot法检测黄芪多糖处理后乳腺癌MDA-MB-231细胞Wnt信号通路Wnt1和β-catenin蛋白的表达;应用ImmPort数据库免疫相关基因集与差异表达基因取交集,进行免疫相关通路分析;考察不同组别程序性细胞死亡配体1(programmed cell death ligand 1,PD-L1)的表达水平。结果 黄芪多糖以浓度依赖性方式抑制乳腺癌细胞增殖作用,与对照组相比,黄芪多糖给药组共筛选出363个差异表达基因,主要富集在Wnt信号通路、ECM受体相互作用、乳腺癌等;Western blot分析进一步证实黄芪多糖能够浓度依赖性抑制Wnt信号通路中Wnt1和β-catenin蛋白的表达。与ImmPort数据库免疫基因集进行交集后获取到42个差异表达的免疫基因,KEGG通路富集结果提示,通路主要富集在抗原提呈、炎症反应和TNF信号通路,提示黄芪多糖可能主要通过增加抗原提呈作用介导其发挥乳腺癌免疫调节作用。黄芪多糖能适度降低PD-L1的表达水平,但差异无统计学意义。结论 黄芪多糖以浓度依赖性方式抑制乳腺癌细胞增殖,主要通过Wnt信号通路发挥其抑制作用,此外免疫调节方面可能通过增加抗原提呈作用介导乳腺癌免疫调节作用。
Objective We aimed to investigate the inhibitory effect of astragalus polysaccharide(APS)on breast cancer cells and predict the participating pathways,verify the mechanism,and enrich and analyze the involved immune pathways.Methods CCK-8 method was used to detect the inhibitory effect of APS(0,0.5,1.0,2.0,4.0,8.0 mg/mL)on proliferation of MDA-MB-231 cells.The gene expression profile of APS administration group and control group was determined,and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis of the screened differentially expressed genes was performed by using R clusterProfiler package.Western blot was used to detect the expression of Wnt1 andβ-catenin of Wnt signaling pathway in breast cancer cells treated with APS.The ImmPort database was utilized to collect immune-related gene sets.Immune-related gene set and differentially expressed genes intersection was analyzed.The association between expression levels of PD-L1 and APS administration were investigated.Results APS inhibits proliferation of breast cancer cells in a concentration-dependent manner.Compared with the control group,a total of 363 differentially expressed genes were screened in astragalus polysaccharide administration group,which mainly enriched in Wnt signaling pathway,ECM receptor interaction and breast cancer signaling pathway,et al.Western blot analysis further confirmed that APS could concentration-dependent inhibit the expression of Wnt1 andβ-catenin proteins in the Wnt signaling pathway.Forty-two differentially expressed immune genes were obtained after intersection with ImmPort database immune gene set,and the enrichment results of KEGG indicated that the pathways were mainly enriched in the antigen-presenting,inflammatory response and TNF signaling pathway,suggesting that APS may play its role in the immune regulation of breast cancer mainly by increasing the antigen-presenting function.APS can moderately reduce the expression level of PD-L1,but the difference is not statistically significant Conclusion APS can inhibit the proliferation of breast cancer cells in a concentration dependent manner,mainly through the Wnt signaling pathway.In addition,APS may mediate the immune regulation of breast cancer by increasing antigen presentation.
作者
谭文超
张艳
TAN Wenchao;ZHANG Yan(Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shengyang 110032,Liaoning,China;Liaoning University of Traditional Chinese Medicine,Shengyang 110847,Liaoning,China)
出处
《辽宁中医药大学学报》
CAS
2024年第3期8-12,共5页
Journal of Liaoning University of Traditional Chinese Medicine
基金
国家自然科学基金项目(81774157)。
关键词
黄芪多糖
基因表达
WNT信号通路
免疫调节
乳腺癌
astragalus polysaccharide
gene expression profile
Wnt signaling pathway
immune regulation
breast cancer
