miRNA-548d-3p represses non-small cell lung cancer growth by perturbing DDX5-mediated pyroptosis through JAK2/STAT3 signaling.

BACKGROUND

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.

METHODS

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.

RESULTS

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.

CONCLUSION

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.