Research Paper Volume 12, Issue 20 pp 20432—20444
Ku70 and Ku80 participate in LPS-induced pro-inflammatory cytokines production in human macrophages and monocytes
- 1 Department of Stomatology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China
- 2 Center of Stomatology, The Second Affiliated Hospital of Soochow University, Suzhou, China
- 3 Department of Orthopaedics, Wujin Hospital Affiliated to Jiangsu University and The Wujin Clinical College of Xuzhou Medical University, Changzhou, China
Received: January 23, 2020 Accepted: July 20, 2020 Published: October 27, 2020https://doi.org/10.18632/aging.103845
How to Cite
Copyright: © 2020 Sun et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
In human macrophages and monocytes, lipopolysaccharide (LPS) induces nuclear factor kappa B (NFκB) activation and pro-inflammatory cytokines production. We tested the possible involvement of Ku70 and Ku80 in the process. In THP-1 macrophages and primary human peripheral blood mononuclear cells (PBMCs), shRNA-induced double knockdown of Ku70 and Ku80 potently inhibited LPS-induced production of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6). Additionally, we developed CRISPR/Cas-9 gene-editing methods to knockout both Ku70 and Ku80 in THP-1 cells and PBMCs. Double knockout (DKO) largely inhibited LPS-induced pro-inflammatory cytokines production. Conversely, in THP-1 cells exogenous overexpression of both Ku70 and Ku80 enhanced the pro-inflammatory cytokines production by LPS. Ku70 and Ku80 co-immunoprecipitated with p65-p52 NFκB complex in the nuclei of LPS-treated THP-1 cells. Significantly, LPS-induced NFκB activation was inhibited by Ku70 plus Ku80 double knockdown or DKO. It was however enhanced with Ku70 and Ku80 overexpression. Together, Ku70 and Ku80 promote LPS-induced NFκB activation and pro-inflammatory response in THP-1 cells and human PBMCs.