基于转录组测序分析芍药苷诱导下口腔黏膜上皮细胞差异表达基因

Transcriptome Sequencing Screened Differentially Expressed Genes in Leuk-1 Cells under Paeoniflorin Condition

目的:基于转录组测序分析芍药苷调控人口腔黏膜上皮细胞基因表达的情况,分析关键基因,探讨芍药苷在口腔黏膜上皮细胞中的作用机制以及其药用价值。方法:实验分为3组,空白对照组、低浓度(5μmol/L)和高浓度(10μmol/L)芍药苷组处理口腔黏膜上皮细胞系Leuk-1,进行转录组测序分析,从而筛选出差异表达基因(DEGs),采用GO和KEGG富集分析对DEGs进行基因功能和途径注释。结果:芍药苷诱导下,Leuk-1细胞中共有1212个显著DEGs;GO富集分析显示,DEGs显著富集在免疫反应、体液免疫应答反应、细胞因子介导的通路反应2’-5’寡腺苷酸合成酶活性、丝氨酸内酞酶活性、细胞外空间、细胞外组分和内质网等功能中;KEGG通路富集显示,DEGs显著富集在单纯疱疹病毒Ⅰ感染、癌症通路、细胞因子与细胞因子受体相互作用、人类乳头瘤病毒感染等信号通路中;采用实时荧光定量PCR技术检测了参与芍药苷诱导的Leuk-1细胞差异表达的基因,其趋势域RNA-seq一致,可以验证RNA-seq技术的准确性。结论:芍药苷在口腔黏膜上皮细胞中的作用主要体现在抗病毒感染、肿瘤抑制、保护上皮完整性以及免疫调节等方面。

Objective:To analyze the gene expression of Leuk-1 cells regulated by paeoniflorin based on transcriptome sequencing,to excavate and analyze key genes,to study the mechanism of paeoniflorin on Leuk-1 cells,and to explore its medicinal value.Methods:Transcriptome analyze were carried out on Leuk-1 cells in zero,low(5μmol/L),and high(10μmol/L)concentration groups.A series of different expression genes(DEGs)were screened,and the gene function and pathways of DEGs were annotated by Gene Ontology enrichment and KEGG enrichment analysis.Results:Among 1212 DEGs identified in Leuk-1 cells under the influence of paeoniflorin,Gene Ontology enrichment analysis revealed significant enrichment in immune response,humoral immune response,cytokine-mediated signaling pathway,2’-5’-oligoadenylate synthetase activity,serine-type endopeptidase activity,extracellular space,extracellular region,endoplasmic reticulum,and other pathways.KEGG pathways enrichment indicated that DEGs were significantly enriched in Herpes simplex virus 1 infection,pathways in cancer,cytokine-cytokine receptor interaction,human papillomavirus infection,and other pathways.RT-qPCR examination of DEGs confirmed that accuracy of RNA-seq analysis.The role of paeoniflorin was mainly reflected in antiviral infection,tumor inhibition,protection of epithelial integrity,and immune regulation.Conclusion:The results of this experiment provided a reference to further study the molecular mechanism and provided clues to explore the prevention and treatment of oral mucosal diseases.