PMID- 36587218 OWN - NLM STAT- MEDLINE DCOM- 20230103 LR - 20230124 IS - 1743-422X (Electronic) IS - 1743-422X (Linking) VI - 19 IP - 1 DP - 2022 Dec 31 TI - Mechanism of autophagy induced by activation of the AMPK/ERK/mTOR signaling pathway after TRIM22-mediated DENV-2 infection of HUVECs. PG - 228 LID - 10.1186/s12985-022-01932-w [doi] LID - 228 AB - BACKGROUND: Dengue virus type 2 (DENV-2) was used to infect primary human umbilical vein endothelial cells (HUVECs) to examine autophagy induced by activation of the adenosine monophosphate-activated protein kinase (AMPK)/extracellular signal-regulated kinase (ERK)/mammalian target of rapamycin (mTOR) signaling pathway following tripartite motif-containing 22 (TRIM22)-mediated DENV-2 infection to further reveal the underlying pathogenic mechanism of DENV-2 infection. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to screen putative interference targets of TRIM22 and determine the knockdown efficiency. The effect of TRIM22 knockdown on HUVEC proliferation was determined using the CCK8 assay. Following TRIM22 knockdown, transmission electron microscopy (TEM) was used to determine the ultrastructure of HUVEC autophagosomes and expression of HUVEC autophagy and AMPK pathway-related genes were measured by qRT-PCR. Moreover, HUVEC autophagy and AMPK pathway-related protein expression levels were determined by western blot analysis. Cell cycle and apoptosis were assessed by flow cytometry (FCM) and the autophagosome structure of the HUVECs was observed by TEM. RESULTS: Western blot results indicated that TRIM22 protein expression levels increased significantly 36 h after DENV-2 infection, which was consistent with the proteomics prediction. The CCK8 assay revealed that HUVEC proliferation was reduced following TRIM22 knockdown (P < 0.001). The TEM results indicated that HUVEC autolysosomes increased and autophagy was inhibited after TRIM22 knockdown. The qRT-PCR results revealed that after TRIM22 knockdown, the expression levels of antithymocyte globulin 7 (ATG7), antithymocyte globulin 5 (ATG5), Beclin1, ERK, and mTOR genes decreased (P < 0.01); however, the expression of AMPK genes (P < 0.05) and P62 genes (P < 0.001) increased. FCM revealed that following TRIM22 knockdown, the percentage of HUVECs in the G2 phase increased (P < 0.001) along with cell apoptosis. The effect of TRIM22 overexpression on HUVEC autophagy induced by DENV-2 infection and AMPK pathways decreased after adding an autophagy inhibitor. CONCLUSIONS: In HUVECs, TRIM22 protein positively regulates autophagy and may affect autophagy through the AMPK/ERK/mTOR signaling pathway. Autophagy is induced by activation of the AMPK/ERK/mTOR signaling pathway following TRIM22-mediated DENV-2 infection of HUVECs. CI - (c) 2022. The Author(s). FAU - Wu, Ning AU - Wu N AD - Chemistry and Biochemistry Laboratory, Guizhou Medical University, Guiyang, China. FAU - Gou, Xiaoqin AU - Gou X AD - Chemistry and Biochemistry Laboratory, Guizhou Medical University, Guiyang, China. FAU - Hu, Pan AU - Hu P AD - Chemistry and Biochemistry Laboratory, Guizhou Medical University, Guiyang, China. FAU - Chen, Yao AU - Chen Y AD - Chemistry and Biochemistry Laboratory, Guizhou Medical University, Guiyang, China. FAU - Ji, Jinzhong AU - Ji J AD - Chemistry and Biochemistry Laboratory, Guizhou Medical University, Guiyang, China. FAU - Wang, Yuanying AU - Wang Y AD - Chemistry and Biochemistry Laboratory, Guizhou Medical University, Guiyang, China. FAU - Zuo, Li AU - Zuo L AD - Department of Immunology, Guizhou Medical University, Guiyang, China. gzykdxzuoli@163.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20221231 PL - England TA - Virol J JT - Virology journal JID - 101231645 RN - EC 2.7.11.31 (AMP-Activated Protein Kinases) RN - EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases) RN - W36ZG6FT64 (Sirolimus) RN - 0 (Antilymphocyte Serum) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - 0 (TRIM22 protein, human) RN - 0 (Tripartite Motif Proteins) RN - 0 (Repressor Proteins) RN - 0 (Minor Histocompatibility Antigens) RN - EC 2.7.1.1 (MTOR protein, human) SB - IM MH - Humans MH - *AMP-Activated Protein Kinases/genetics/metabolism MH - Human Umbilical Vein Endothelial Cells MH - *Extracellular Signal-Regulated MAP Kinases/metabolism MH - Sirolimus/pharmacology MH - Antilymphocyte Serum/pharmacology MH - Signal Transduction MH - TOR Serine-Threonine Kinases/genetics/metabolism/pharmacology MH - Autophagy MH - Tripartite Motif Proteins/genetics/pharmacology MH - Repressor Proteins/metabolism MH - Minor Histocompatibility Antigens/pharmacology PMC - PMC9805691 OTO - NOTNLM OT - AMPK/ERK/mTOR signaling pathway OT - Autophagy OT - DENV-2 OT - TRIM22 COIS- The authors declare that they have no competing interests. EDAT- 2023/01/01 06:00 MHDA- 2023/01/04 06:00 PMCR- 2022/12/31 CRDT- 2022/12/31 23:56 PHST- 2022/06/01 00:00 [received] PHST- 2022/11/22 00:00 [accepted] PHST- 2022/12/31 23:56 [entrez] PHST- 2023/01/01 06:00 [pubmed] PHST- 2023/01/04 06:00 [medline] PHST- 2022/12/31 00:00 [pmc-release] AID - 10.1186/s12985-022-01932-w [pii] AID - 1932 [pii] AID - 10.1186/s12985-022-01932-w [doi] PST - epublish SO - Virol J. 2022 Dec 31;19(1):228. doi: 10.1186/s12985-022-01932-w.