IL-1β通过STAT3信号通路上调食管鳞癌细胞PD-L1表达的研究

IL-1βup-regulates PD-L1 through STAT3 signaling pathway in esophageal squamous cell carcinoma

目的探讨促炎细胞因子白介素1β(IL-1β)对食管鳞癌细胞程序性细胞死亡1配体1(PD-L1)的表达调控作用及调控机制,阐明IL-1β在食管鳞癌肿瘤微环境中调控PD-L1表达的临床意义。方法分析TCGA数据中收录的有关食管癌中IL-1β和PD-L1表达的数据,明确IL-1β和PD-L1在食管癌中的表达及预后意义;在体外细胞系水平,运用重组的人源化IL-β蛋白刺激食管鳞癌细胞系KYSE30后,运用qRT-PCR检测PD-L1的mRNA表达,运用免疫印迹和免疫荧光实验技术检测PD-L1蛋白表达;为了明确IL-β的作用信号通路,运用抗体芯片筛选重组的人源化IL-β蛋白处理和未处理食管癌细胞内的信号通路差异。结果生物信息学分析显示,相比癌旁正常组织IL-1β和PD-L1在食管癌组织中显著性上调表达;但上调的IL-1β和PD-L1表达与食管癌患者的总生存差具有统计学趋势但不具有统计学相关性。重组的人源化IL-β蛋白刺激食管鳞癌细胞系后,能显著性上调PD-L1的mRNA和蛋白表达。抗体芯片筛选结果显示,重组的人源化IL-β蛋白处理食管癌细胞后,能显著性激活STAT3信号通路。结论IL-1β通过STAT3信号通路上调食管鳞癌细胞PD-L1表达。

Objective To explore the underlying mechanism by which Interleukin1β(IL-1β)regulates over Programmed Death Ligand-1(PD-L1)in esophageal squamous cell carcinoma(ESCC)and to understand the prognostic meaning of IL-1βand PD-L1 expressions in ESCC.Methods To understand the expression status and its prognostic significance of IL-1βand PD-L1 expressions in esophageal carcinoma,data were bioinformatically analyzed with GEPIA and TIMER databases;In vitro cell culture system,variation of mRNA of PD-L1 was detected using quantitative real-time PCR(qRT-PCR)in ESCC cell line KYSE30 treated with or without recombinant human IL-1βproteins;Likewise,the protein variation of PD-L1 was evaluated by western-blot and immunofluorescent staining,respectively.To determine which signaling pathway got involved in the treatment of IL-1β,antibody microarray was employed.Results Bioinformatically analyzed with GEPIA and TIMER databases,IL-1βand PD-L1 were shown to be significantly elevated in esophageal carcinoma compared with normal control;prognostically,there was a trend towards significance of IL-1βand PD-L1 expression in esophageal carcinoma;however,no significance can be achieved.Treated with recombinant human IL-1βprotein,KYSE30 cells whose PD-L1 can be markedly up-regulated on both mRNA and protein levels.Data from antibody microarray showed that it was STAT3 signaling pathway that got involved in the regulation of IL-1βover PD-L1 in ESCC cells.Conclusion IL-1βup-regulates PD-L1 through STAT3 signaling pathway in esophageal squamous cell carcinoma.