STAT3诱捕寡核苷酸经STAT3/IRF-1通路抑制肿瘤细胞PD-L1的表达  

Inhibition of PD-L1 expression with STAT3 decoy oligonucleotide in tumor cells via STAT3/IRF-1 pathway

作　　者：依含 黄建胜[1] 李雪 朱乃硕[1] YI Han;HUANG Jian-Sheng;LI Xue;ZHU Nai-Shuo(College of Life Science,Fudan University,Shanghai 200438,China)

机构地区：[1]复旦大学生命科学学院

出　　处：《中国免疫学杂志》2019年第22期2742-2748,2754,共8页Chinese Journal of Immunology

基　　金：国家科技重大专项(2012ZX10002006-002-003);国家自然科学基金(30571650,31370927);上海市科委科学基金(13431900602)资助项目

摘　　要：目的:探究STAT3诱捕寡核苷酸(STAT3 Decoy-ON)对免疫抑制分子PD-L1的调控及其作用机制。方法:以前列腺癌细胞DU145为模型,通过流式细胞仪与激光扫描共聚焦显微镜分析STAT3 Decoy-ON对DU145的亲和性;通过qRT-PCR与Western blot分析STAT3 Decoy-ON对STAT3与IRF-1的表达影响;通过流式细胞仪及上述方法分析STAT3 Decoy-ON对PD-L1的表达调控;进一步构建PD-L1启动子上STAT3、IRF-1转录因子识别位点的突变或缺失荧光素酶质粒,探究STAT3与IRF-1对PD-L1表达的调控。结果:发现STAT3 Decoy-ON与DU145细胞具有高度亲和性,且显著下调DU145细胞STAT3与PD-L1的表达;同时STAT3与IRF-1识别位点的缺失或突变皆可显著降低PD-L1启动子的活性;STAT3 Decoy-ON通过下调细胞内IRF-1的表达发挥对PD-L1的抑制作用。结论:STAT3 Decoy-ON介导STAT3/IRF-1的信号通路抑制前列腺癌细胞(DU145)PD-L1的表达。Objective:To study the regulation and mechanism of PD-L1 expression with STAT3 decoy oligonucleotide in tumor cells.Methods:The prostate cancer cell DU145 was used as research model,the affinity between STAT3 decoy oligonucleotide and tumor cells was displayed by flow cytometry and laser scanning confocal microscopy.The STAT3 and IRF-1 expression level in tumor cells was analyzed by qRT-PCR and Western blot.Then the PD-L1 expression level was analyzed by flow cytometry,qRT-PCR and Western blot.The mutant or deficient luciferase plasmid of STAT3 and IRF-1 transcription factor recognition sites on the PD-L1 promoter was further constructed to explore the regulatory mechanism of PD-L1 expression.Results:STAT3 Decoy-ON had a high affinity with DU145,and down-regulated the expression of STAT3 and PD-L1 in cells significantly.The mutation or deficiency of STAT3 and IRF-1 binding sites could inhibit the activity of PD-L1 promoter significantly,and STAT3 Decoy-ON could play an inhibitory role on PD-L1 by down-regulating the expression of IRF-1 in DU145.Conclusion:STAT3 Decoy-ON inhibits the expression of PD-L1 in prostate cancer cells(DU145)by regulating the signal pathway of STAT3/IRF-1.

关 键 词：STAT3诱捕寡核苷酸 PD-L1 抗肿瘤药物 

分 类 号：R730.51[医药卫生—肿瘤]