摘要
目的 本研究旨在系统分析Toll样受体7(TLR7)基因在胶质母细胞瘤(GBM)中的表达特征,并探索其在GBM中的功能及上游调控机制。方法 TIMER 2.0和UALCAN数据库分析TLR7在GBM和癌旁组织中的表达情况。利用miRWalk、TargetScanHuman 8.0、Tarbase、miRDB和miRTarBase数据库预测TLR7 3'UTR区可能结合的miRNA。通过TRANSFAC和ContraV3数据库分析TLR7基因启动子区潜在结合的转录因子。使用双荧光素酶报告基因实验验证hsa-miR-758-3p和核因子κB(NF-κB)与TLR7的靶向调控关系。结果 TLR7在GBM中的表达显著高于癌旁正常组织(P<0.001),且与患者的年龄、肿瘤蛋白P53(TP53)突变状态和性别密切相关。TLR7在胆管癌、胶质母细胞瘤、肾透明细胞癌等七种肿瘤中高表达,而在结肠癌、肺腺癌、肺鳞癌等七种癌症中低表达(P<0.05)。生物信息学预测发现,hsa-miR-758-3p可能结合于TLR7的3'UTR,转录因子NF-κB结合于TLR7的启动子区。双荧光素酶报告基因实验结果显示,miR-758-3p可导致TLR7 3'UTR在U87和U251细胞中的转录活性分别降低32%和23%(P<0.01);突变TLR7的miR-758-3p结合位点后,其转录活性不受mi R-758-3p影响。此外,TLR7在U87、U251细胞中的启动子活性受NF-κB抑制剂影响,分别降低70%和55%(P<0.001);当突变NF-κB结合位点后,荧光素酶报告基因活性不受NF-κB抑制剂影响。结论 本研究揭示了TLR7在GBM中的表达特征及其上游调控机制,为进一步理解TLR7在GBM中的功能及其作为潜在治疗靶点的可能性提供了重要的理论依据。
Objective The aim of this study was to systematically analyze the expression characteristics of Toll-like receptor 7(TLR7)gene in glioblastoma(GBM),and explore its function and upstream regulatory mechanism in GBM.Method The expression of TLR7 in GBM and para-cancerous tissues was analyzed by TIMER 2.0 and UALCAN database.miRWalk,TargetScanHuman 8.0,Tarbase,miRDB and miRTarBase databases were used to predict the potential binding mirnas of TLR73'UTR region.TRANSFAC and ContraV3 databases were used to analyze potential binding transcription factors of TLR7 gene promoter region.Dual luciferase reporter gene assay was used to verify the targeted regulatory relationship between hsa-miR-758-3p and nuclear factorκB(NF-κB)and TLR7.Results The expression of TLR7 in GBM was significantly higher than that in para-cancer normal tissues(P<0.001),and was closely related to the age,mutation status of tumor protein P53(TP53)and gender of the patients.TLR7 was highly expressed in seven cancers,including cholangiocarcinoma,glioblastoma and renal clear cell carcinoma,but was low expressed in seven cancers,including colon cancer,lung adenocarcinoma and lung squamous carcinoma(P<0.05).Bioinformatics predicted that hsa-miR-758-3p may bind to the 3’utr of TLR7,and the transcription factor NF-κB may bind to the promoter region of TLR7.The results of double luciferase reporter gene assay showed that miR-758-3p could reduce the transcriptional activity of TLR73'UTR in U87 and U251 cells by 32%and 23%,respectively(P<0.01);After mutating the miR-758-3p binding site of TLR7,its transcriptional activity was not affected by miR-758-3p.In addition,the promoter activity of TLR7 in U87 and U251 cells was reduced by 70%and 55%by NF-κB inhibitors,respectively(P<0.001).When the NF-κB binding site was mutated,luciferase reporter gene activity was not affected by NF-κB inhibitors.Conclusion This study revealed the expression characteristics of TLR7 in GBM and its upstream regulatory mechanism,providing an important theoretical basis for further understanding the function of TLR7 in GBM and its possibility as a potential therapeutic target.
作者
郭力雯
周先雷
仵红娇
徐鸿学
张洪梅
吴文龙
张雪梅
GUO Liwen;ZHOU Xianlei;WU Hongjiao;XU Hongxue;ZHANG Hongmei;WU Wenlong;ZHANG Xuemei(College of Life Science,North China University of Science and Technology,Tangshan 063210,China)
出处
《中国煤炭工业医学杂志》
2024年第5期447-453,共7页
Chinese Journal of Coal Industry Medicine
基金
2021年度唐山市人才项目(编号:A2021100007)
