Wu Chen, Li Xiaodong, Zhang Dachuan, Xu Bin, Hu Wenwei, Zheng Xiao, Zhu Danxia, Zhou Qi, Jiang Jingting, Wu Changping
Department of Oncology, the Third Affiliated Hospital of Soochow University, Changzhou, China.
Institute of Cell Therapy, Soochow University, Changzhou, China.
Cell Physiol Biochem. 2018;48(6):2493-2502. doi: 10.1159/000492687. Epub 2018 Aug 17.
BACKGROUND/AIMS: IL-1β is an important mediator of "inflammation-cancer" transformation through IL-1β/NF-κB/COX-2/HIF-1α signaling pathway, whereas certain portion of patients with lung adenocarcinoma (LUAD) still suffer from rapid tumor progression in clinical practice, indicating the occurrence of potential bypass.
Real-time polymerase chain reaction was applied to examine the expressions of mir-144-3p, WT1, NF-κB, COX2 and HIF-1α at the mRNA level in 127 LUAD samples and corresponding adjacent tissues. miR-144-3p mimic and antagormiR were used to trigger activation and suppression of miR-144-3p in A549 cells, respectively. MTT assay and Western blotting analysis were carried out to evaluate the cell proliferation. Stable clones with over-expression or knockdown of WT1 were generated with plasmid or shRNA by lentiviral vector technology in H1568 and H1650 NSCLC cell lines, respectively. Dual luciferase reporter assay was performed to validate the effect of miR-144-3p on WT1D. Xenograft model was established for in vivo experiment, and TCGA data were extracted for validation.
miR-144-3p could suppress the WT1D expression at the post-transcriptional level, hence regulating cell proliferation in LUAD. WT1 and COX-2 were independent prognostic factors of LUAD patients. In addition, inhibition of IL-1β/miR-144-3p/WT1D and IL-1β/NF-κB/COX-2/HIF-1α pathways using miR-144-3p mimic and Celecoxib, respectively, displayed synergistic suppressive effect on cell proliferation in LUAD.
A de novo IL-1β/miR-144-3p/WT1D axis was involved in proliferative regulation of LUAD. Moreover, simultaneous blockade of both IL-1β/miR-144-3p/WT1D and IL-1β/NF-κB/COX-2/ HIF-1α pathways might have synergistic suppressive effect on cell proliferation in LUAD.
背景/目的：白细胞介素-1β（IL-1β）是通过IL-1β/核因子κB（NF-κB）/环氧化酶-2（COX-2）/缺氧诱导因子-1α（HIF-1α）信号通路介导“炎症-癌症”转化的重要介质，然而在临床实践中，仍有部分肺腺癌（LUAD）患者肿瘤进展迅速，提示可能存在潜在的旁路途径。
应用实时聚合酶链反应检测127例LUAD样本及相应癌旁组织中mir-144-3p、威尔姆斯瘤基因1（WT1）、NF-κB、COX2和HIF-1α的mRNA水平表达。分别用miR-144-3p模拟物和抗miR-144-3p在A549细胞中激活和抑制miR-144-3p。采用MTT法和蛋白质印迹分析评估细胞增殖情况。分别通过慢病毒载体技术利用质粒或短发夹RNA（shRNA）在H1568和H1650非小细胞肺癌（NSCLC）细胞系中构建WT1过表达或敲低的稳定克隆。进行双荧光素酶报告基因检测以验证miR-144-3p对WT1D的作用。建立异种移植模型进行体内实验，并提取癌症基因组图谱（TCGA）数据进行验证。
miR-144-3p可在转录后水平抑制WT1D表达，从而调节LUAD细胞增殖。WT1和COX-2是LUAD患者的独立预后因素。此外，分别使用miR-144-3p模拟物和塞来昔布抑制IL-1β/miR-144-3p/WT1D和IL-1β/NF-κB/COX-2/HIF-1α通路，对LUAD细胞增殖显示出协同抑制作用。
一条全新的IL-1β/miR-144-3p/WT1D轴参与LUAD的增殖调控。此外，同时阻断IL-1β/miR-144-3p/WT1D和IL-1β/NF-κB/COX-2/HIF-1α两条通路可能对LUAD细胞增殖具有协同抑制作用。
