Liu Liyu, Liu Qinglin, Wu Zhining, Wu Jiao, Chen Xiaoyan, Zhang Bocheng, Pang Xiao, Liang Shixuan, Long Ying, Liu Ying
Department of Thoracic Medicine, The Affiliated Cancer Hospital of Xiangya School of Medicine/Hunan Cancer Hospital, Central South University, Changsha, Hunan 410013, PR China.
Department of Clinical Laboratory, The Affiliated Cancer Hospital of Xiangya School of Medicine/Hunan Cancer Hospital, Central South University, Changsha, Hunan 410013, PR China.
Cell Signal. 2025 Jun 15;134:111945. doi: 10.1016/j.cellsig.2025.111945.
The function of microRNAs (miRNAs) in tumor development has been extensively characterized. Pyroptosis is a kind of programmed death, which can effectively hinder cancer development. However, there is still a large gap in the function of miRNAs in NSCLC pyroptosis.
The pyroptosis-related differentially expressed miRNAs and their promising targets in NSCLC were analyzed using bioinformatic analyses. The effects of miR-548d-3p on cell proliferation, pyroptosis and tumor growth were verified in vivo and in vitro. The expression of pyroptosis-related factors was examined in cells and xenografted tumors. Additionally, the molecular interaction was assessed by using Co-IP and dual-luciferase reporter gene assays.
We found that miR-548d-3p was poorly expressed in NSCLC in public datasets and an independent cohort of 48 NSCLC patients. Low miR-548d-3p expression was positively associated with pathologic T stage and poor prognosis of NSCLC patients. Transfection of miR-548d-3p mimics significantly decreased the cell viability of NSCLC cells, partly attributing to the increase in the proportion of pyroptotic cells. These changes were accompanied by a rise in the protein abundance of NLRP3, ASC, cleaved Caspase-1 and GSDMD-N and release of IL-1β and IL-18. Integrating bioinformatic, expressional and experimental analyses, we predicted and validated DDX5 as the direct target of miR-548d-3p. Furthermore, we demonstrated that miR-548d-3p/DDX5 axis regulated pyroptosis via the Phosphorylated JAK2/STAT3-mediated NLRP3/Caspase-1/GSDMD pathway in vitro and in vivo.
Our study revealed that miR-548d-3p/DDX5 promoted pyroptosis in NSCLC by promoting the JAK2/STAT3/NLRP3/Caspase-1/GSDMD pathway, indicating the promising effect of miR-548d-3p in NSCLC treatment.
微小RNA（miRNA）在肿瘤发生发展中的作用已得到广泛研究。细胞焦亡是一种程序性死亡，可有效抑制癌症发展。然而，miRNA在非小细胞肺癌（NSCLC）细胞焦亡中的作用仍存在很大差距。
利用生物信息学分析方法分析NSCLC中与细胞焦亡相关的差异表达miRNA及其潜在靶点。在体内和体外验证了miR-548d-3p对细胞增殖、细胞焦亡和肿瘤生长的影响。检测细胞和异种移植瘤中细胞焦亡相关因子的表达。此外，通过免疫共沉淀（Co-IP）和双荧光素酶报告基因实验评估分子间相互作用。
我们发现，在公开数据集以及48例NSCLC患者的独立队列中，miR-548d-3p在NSCLC中表达较低。miR-548d-3p低表达与NSCLC患者的病理T分期及不良预后呈正相关。转染miR-548d-3p模拟物可显著降低NSCLC细胞的活力，部分原因是细胞焦亡比例增加。这些变化伴随着NLRP3、ASC、裂解的半胱天冬酶-1和GSDMD-N蛋白丰度的升高以及IL-1β和IL-18的释放。综合生物信息学、表达和实验分析，我们预测并验证了DDX5是miR-548d-3p的直接靶点。此外，我们证明miR-548d-3p/DDX5轴在体内外通过磷酸化的JAK2/STAT3介导的NLRP3/半胱天冬酶-1/GSDMD途径调节细胞焦亡。
我们研究表明，miR-548d-3p/DDX5通过促进JAK2/STAT3/NLRP3/半胱天冬酶-1/GSDMD途径促进NSCLC细胞焦亡，提示miR-548d-3p在NSCLC治疗中具有潜在疗效。
