Effect of LncRNA XIST on apoptosis of bronchial epithelial cells infected with respiratory syncytial virus by regulating the miR-574-5p/TLR4 axis
FU Xiaokang
LIU Baojuan
GAO Shuai
SU Qin
WEI Quanping
SUN Zimei
Abstract：Objective To investigate the effect of long non-coding (Lnc) ribonucleic acid (RNA) X-inactive specific transcript (XIST) on apoptosis of bronchial epithelial cells infected with respiratory syncytial virus (RSV) by regulating the microRNA (miR)-574-5p/Toll-like receptor 4 (TLR4) axis. Methods Human bronchial epithelial cells (16HBE) were separated into negative control (NC) group,RSV infection (RSV) group,small interfering RNA negative control (si-NC) group,XIST small interfering RNA (si-XIST) group,si-XIST+inhibitor negative control (inhibitor NC) group,and si-XIST+miR-574-5p inhibitor group,Real-time fluorescence quantitative polymerase chain reaction was applied to detect the expression of LncRNA XIST and miR-574-5p;5-ethynyl-2'-deoxyuridine (EdU) staining and flow cytometry were applied to detect cell proliferation and apoptosis,respectively;enzyme linked immunosorbent assay was applied to detect the secretions of tumor necrosis factor (TNF)-α,interleukin (IL)-6,IL-1β;dual luciferase reporter gene experiment was applied to verify the relationship between miR-574-5p and LncRNA XIST,TLR4. Western blot was applied to detect the expression of TLR4,proliferating cell nuclear antigen (PCNA),anti apoptotic factor B cell lymphomatoma-2 (Bcl-2),Bcl-2 associated X protein (Bax),and cleaved cysteine aspartate proteolytic enzyme 3 (Cleaved caspase-3) proteins. Results Compared with the NC group,the level of LncRNA XIST,apoptosis rate,the level of IL-6,IL-1β,TNF-α,the expression of TLR4,Bax,and Cleaved Caspase-3 in RSV group and si-NC group were elevated,the positive rate of EdU,the level of miR-574-5p,the expression of PCNA and Bcl-2 were decreased (P<0.05);compared with the si-NC group,the level of LncRNA XIST,apoptosis rate,the level of IL-6,IL-1β,TNF-α,the expression of TLR4,Bax,and Cleaved Caspase-3 in the si-XIST group were decreased,the positive rate of EdU,the level of miR-574-5p,the expression of PCNA and Bcl-2 were increased (P<0.05);downregulation of miR-574-5p was able to reverse the inhibitory effects of knocking down LncRNA XIST on apoptosis and inflammatory response of 16HBE cells infected with RSV (P<0.05);dual luciferase reporter gene experiment confirmed the targeted regulatory relationship between miR-574-5p and LncRNA XIST,and between miR-574-5p and TLR4 (P<0.05). Conclusion LncRNA XIST is upregulated in 16HBE cells infected with RSV,and knocking down LncRNA XIST may down-regulate the expression of TLR4 by up-regulating miR-574-5p,and inhibit apoptosis of 16HBE cells after RSV infection.
Keywords：long non-coding ribonucleic acid X-inactive specific transcriptmicroRNA-574-5p/Toll-like receptor axisrespiratory syncytial virusbronchial epithelial cellsapoptosis
Publication Date：2024-08-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 345-351 )