河北医科大学学报 ›› 2023, Vol. 44 ›› Issue (7): 814-818,829.doi: 10.3969/j.issn.1007-3205.2023.07.013

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STING在膀胱癌患者组织表达及对IFN-γ/STAT1信号通路磷酸化修饰调控的机制研究

  

  1. 1.南京医科大学附属无锡人民医院泌尿外科，江苏 无锡 214023； 2.南京医科大学附属无锡人民医院超声医学科，江苏 无锡 214023

  • 出版日期:2023-07-25 发布日期:2023-07-24
  • 作者简介:王勇 (1984-)，男，江苏无锡人，南京医科大学附属无锡人民医院副主任医师，医学硕士，从事泌尿系肿瘤诊治研究。
  • 基金资助:
    无锡市人才双百后备拔尖人才（HB2020001）

Expression of STING in bladder cancer tissues and its mechanism of regulating phosphorylation modification of IFN- γ/STAT1 signaling pathway

  1. 1.Department of Urology, Wuxi People′s Hospital Affiliated to Nanjing Medical University, Jiangsu 
    Province, Wuxi 214023， China; 2.Department of Ultrasound, Wuxi People′s Hospital Affiliated to 
    Nanjing Medical University, Jiangsu Province, Wuxi 214023， China

  • Online:2023-07-25 Published:2023-07-24

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摘要： 目的 探究干扰素基因刺激蛋白（stimulator of interferon genes， STING）在膀胱癌患者中的表达及对干扰素信号通路干扰素γ（interferon-γ，IFN-γ）/信号传导及转录激活蛋白1(signal transducer and activator of transcription1，STAT1）的调控机制研究。
方法 队列试验：选取膀胱癌患者110例，收集癌组织及癌旁组织样本，通过实时荧光定量聚合酶链式反应(quantitative realtime polymerase chain reaction,qPCR)法检测癌组织和癌旁组织STING表达水平。细胞学试验：采用CRISPR-Cas9技术敲除人膀胱癌BIU-87细胞STING基因，将膀胱癌细胞为3组，对照组、NC组（转染NC质粒）和试验组（转染CRISPR-Cas9质粒），MTT法检测在24 h，48 h,72 h不同时间点细胞的增殖能力，通过流式细胞技术检测各组细胞存活、早期凋亡和晚期凋亡比例，qPCR 检测IFN-γ和STAT1 mRNA 表达水平，Western blot法检测 IFN-γ和STAT1 蛋白表达水平，检测STAT1蛋白磷酸化修饰水平。
结果 队列试验：膀胱癌组织STING表达水平高于癌旁组织（P＜0.05）。细胞学试验：与对照组比较，敲除STING组细胞增殖能力降低，并且随着时间的延长增殖能力进一步降低（P＜0.05）；并且细胞存活降低，细胞早期凋亡和晚期凋亡比例增加（P＜0.05）；IFN-γ和STAT1 mRNA表达水平降低（P＜0.05）；IFN-γ和STAT1蛋白表达水平降低，并且STAT1磷酸化水平降低（P＜0.05）。
结论  STING可能通过激活干扰素信号通路IFN-γ/STAT1的蛋白表达及磷酸化修饰参与膀胱癌的发生发展过程。


关键词: 膀胱肿瘤, 干扰素基因刺激蛋白, 干扰素γ

Abstract: Objective To investigate the expression of stimulator of interferon genes (STING) in patients with bladder cancer and its mechanism of regulating interferon- γ (IFN-γ)/signal transducer and activator of transcription 1 (STAT1) signaling pathway. 
Methods In this cohort study, 110 patients with bladder cancer were selected to collect cancer tissues and adjacent tissues, and the expression of STING in cancer tissues and adjacent tissues was detected by quantitative real-time polymerase chain reaction (qPCR). In cytological test, CRISPR-Cas9 technology was used to knock out the STING gene of human bladder cancer BIU-87 cells, and bladder cancer cells were divided into three groups: control group, NC group (transfected with NC plasmid) and experimental group (transfected with CRISPR-Cas9 plasmid). MTT method was used to detect the proliferation ability of cells at different time points (at 24 h, 48 h and 72 h), and flow cytometry was used to detect the proportion of cell survival, early apoptosis and late apoptosis in each group. qPCR was used to detect mRNA expression levels of IFN- γ and STAT1, and Western blot method for detecting IFN- γ and STAT1 protein expression level as well as the phosphorylation modification level of STAT1 protein. 
Results In cohort study, the expression level of STING in bladder cancer tissue was higher than that in adjacent tissue (P＜0.05). Cytological test showed that, compared with the control group, the proliferation ability of cells in the knockout STING group decreased, and further decreased with time (P＜0.05). Cell proliferation decreased cell survival, and the proportion of early and late apoptosis increased (P＜0.05). IFN- γ and STAT1 mRNA expression levels decreased (P＜0.05), and the expression level of IFN- γ and STAT1 protein as well as the phosphorylation level of STAT1 decreased (P＜0.05). 
Conclusion STING may activate the protein expression of IFN-γ/STAT1 signaling pathway and phosphorylation modification of STAT1, thereby getting involved in the occurrence and development of bladder cancer. 


Key words: urinary bladder neoplasms, stimulator of interferon genes, interferon-γ 

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