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Effects of let‐7e on LPS‐Stimulated THP‐1 Cells Assessed by iTRAQ Proteomic Analysis
Author(s) -
Gui Lian,
Zhang Qianqian,
Cai Yan,
Deng Xiaohong,
Zhang Yingke,
Li Cheukfai,
Guo Qi,
He Xiaoshun,
Huang Junqi
Publication year - 2018
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201700012
Subject(s) - thp1 cell line , computational biology , microbiology and biotechnology , chemistry , biology , cell culture , genetics
Purpose Previous studies have demonstrated that let‐7e is associated with inflammatory responses. To date, the roles and mechanisms of let‐7e have not been completely revealed.Therefore, we aim to identify proteins associated with let‐7e overexpression and explore their functions in the immune responses, including in cytokine production. Experimental design High‐throughput isobaric tag for relative and absolute quantitation (iTRAQ) technology is used to provide the first genome‐wide study of THP‐1 cells transfected with let‐7e mimic followed by lipopolysaccharide (LPS) stimulation. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database and KEGG pathway enrichment analyses are used to analyze a large number of differentially expressed proteins (DEPs) associated with let‐7e overexpression or LPS stimulation. Quantitative reverse transcription PCR (qRT‐PCR) and 50% tissue culture infective dose (TCID50) assays are processed to confirm the relationship of let‐7e and dengue virus replication. Results iTRAQ results show that let‐7e is associated with the expression of anti‐viral proteins. What's more, calcineurin subunit B type 1, an anti‐tumor factor, is upregulated by let‐7e after LPS stimulation. KEGG analyses identify that some DEPS associated with let‐7e overexpression are involved in the measles and influenza A pathways, and LPS‐stimulated proteins in THP‐1 cells are mainly enriched in transcriptional misregulation in cancer pathway and hippo signaling pathway (multiple species). The results of qRT‐PCRand TCID50 show that let‐7e promotes dengue virus replication, which is in agreement with the iTRAQ results. Conclusions and clinical relevance These results provide molecular insights into the regulatory mechanisms of let‐7e in cytokine expression, virus replication, and anti‐tumor function.