摘要
目的:探讨白细胞介素-17(interleukin-17,IL-17)对人肝内胆管癌细胞程序性细胞死亡蛋白配体1(programmed cell death ligand 1,PD-L1)表达水平的影响及其潜在机制。方法:使用不同浓度(0 ng/mL、5 ng/mL、10 ng/mL、20 ng/mL、50 ng/mL和100 ng/mL)的IL-17作用于人肝内胆管癌细胞系RBE和HCCC9810,Western blot检测PD-L1的表达情况,并筛选出IL-17的最佳作用浓度。使用最佳作用浓度处理RBE和HCCC9810细胞,收集mRNA、蛋白质。实时定量逆转录-聚合酶链反应(qRT-PCR)检测PD-L1在mRNA水平的表达情况。Western blot检测JAK2/STAT3信号通路的活化情况,并使用AG490阻断信号通路,Westernblot检测通路被阻断后,IL-17对RBE及HCCC9810细胞PD-L1表达水平的影响。结果:IL-17诱导了PD-L1在RBE和HCCC9810细胞中的表达,并且PD-L1的上调幅度与IL-17的浓度相关,当浓度为50 ng/mL和100 ng/mL时,对PD-L1的诱导作用最为显著。与对照组相比,50 ng/mL的IL-17可显著刺激PD-L1在mRNA水平的表达(P<0.01)。同对照组相比,50 ng/mL的IL-17促进了STAT3和JAK2蛋白的磷酸化水平(P<0.01),但并不影响STAT3和JAK2总蛋白的表达水平(P>0.05)。在使用AG490阻断JAK2/STAT3信号通路后,IL-17诱导PD-L1蛋白表达的能力明显减弱(P<0.05)。结论:IL-17可诱导PD-L1在人肝内胆管癌细胞系RBE和HCCC9810中的表达,其机制可能与促进JAK2/STAT3信号通路蛋白的磷酸化水平有关。
Objective:To investigate the effect of interleukin-17(IL-17)on programmed cell death ligand 1(PD-L1)expression level and the mechanism in intrahepatic cholangiocarcinoma cells.Methods:Human intrahepatic cholangiocarcinoma cell lines RBE and HCCC9810 were cultured in vitro and stimulated with different concentrations(0 ng/mL,5 ng/mL,10 ng/mL,20 ng/mL,50 ng/mL and 100 ng/mL)of IL-17,respectively,and the expression of PD-L1 was detected by Western blot,and the optimal concentration of drug action was determined.After the treatment of RBE and HCCC9810 with the optimal drug concentration,mRNA and protein were collected.Quantitative real-time reverse transcription-polymerase chain reaction(qRT-PCR)was applied to detect the expression of PD-L1 at the mRNA level.Western blot was used to detect the activation of JAK2/STAT3 signaling pathway,and the signaling pathway was blocked using related signaling pathway inhibitor AG490 according to the results,and Western blot was used to detect the expression of PD-L1 in two cell lines(RBE and HCCC9810)after the signaling pathway was blocked.Results:IL-17 significantly upregulated PD-L1 expression in RBE cell line and HCCC9810 cell line.The up-regulation of PD-L1 was related to the concentration of IL-17,with the most significant upregulation effect at concentrations of 50 ng/mL and 100 ng/mL.Compared with IL-17 at 0 ng/mL,IL-17 at 50 ng/mL significantly stimulated PD-L1 expression at the mRNA level(P<0.01).Compared with the control group,IL-17 at 50 ng/mL significantly up-regulated the phosphorylation levels of STAT3 protein and JAK2 protein in RBE cell line and HCCC9810 cell line(P<0.01),but did not affect the levels of total STAT3 protein and JAK2 protein(P>0.05).After blocking the JAK2/STAT3 signaling pathway by AG490,the ability of IL-17 to induce the upregulation of PD-L1 protein level was significantly weakened(P<0.05).Conclusion:IL-17 could upregulate PD-L1 expression in human intrahepatic cholangiocarcinoma cell lines RBE and HCCC9810,and the mechanism may be related to th
作者
段家康
张懿刚
庞青
范龙飞
刘双池
尹宏祥
王蕊
谈燚
DUAN Jiakang;ZHANG Yigang;PANG Qing;FAN Longfei;LIU Shuangchi;YIN Hongxiang;WANG Rui;TAN Yi(Department of Hepatobiliary Surgery,the First Affiliated Hospital of Bengbu Medical College,Anhui Bengbu 233004,China;Bengbu Medical College,Anhui Bengbu 233030,China)
出处
《现代肿瘤医学》
CAS
北大核心
2021年第24期4265-4270,共6页
Journal of Modern Oncology
基金
国家自然科学基金(编号:81600452)
安徽高校自然科学研究重大项目(编号:KJ2018ZD022)
蚌埠医学院大学生创新创业课题(编号:S202010367075,S202010367004)。
