TY - JOUR
T1 - Effects of let-7e on LPS-Stimulated THP-1 Cells Assessed by iTRAQ Proteomic Analysis
AU - Gui, Lian
AU - Zhang, Qianqian
AU - Cai, Yan
AU - Deng, Xiaohong
AU - Zhang, Yingke
AU - Li, Cheukfai
AU - Guo, Qi
AU - He, Xiaoshun
AU - Huang, Junqi
N1 - Funding Information:
Conceived and designed the experiments: J.H., Y.Z., L.G. Performed the experiments: Y.Z., L.G. Analyzed the data: L.G., Q.Z., Y.C., Q.G., X.H. Contributed reagents/materials/analysis tools: J.H., L.G., Q.Z., Y.C., C.L., X.D., Q.G. Wrote the manuscript: L.G., Q.Z. The present study was supported by National Natural Science Foundation of China (No. 31370870 and No. 30872350), Natural Science Foundation of Guangdong Province (No. s81510008901000017, No. s2012010009050, and No. s2013020013000), Guangdong Province Scientific Technology Project (No. 2010B050700008, No. 2011B040300022, No. 2013A020229003, and 2015B050501002), Guangzhou City Scientific Technology Project (No. 201604020083, No. 11C32060749, No. 2011J4100084, and No. 2008Z1-E221), The Fundamental Research Funds for the Central Universities (No. 10YKPY31), Guangdong Provincial Key Laboratory Construction Projection on Organ Donation and Transplant Immunology (No. 2013A061401007), and Guangdong Provincial international Cooperation Base of Science and Technology (Organ Transplantation; No. 2015B050501002).
Publisher Copyright:
© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2018/9
Y1 - 2018/9
N2 - 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.
AB - 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.
KW - Cytokines
KW - Proteomic
KW - Virus
KW - let-7e
UR - http://www.scopus.com/inward/record.url?scp=85044751853&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85044751853&partnerID=8YFLogxK
U2 - 10.1002/prca.201700012
DO - 10.1002/prca.201700012
M3 - Article
C2 - 29505169
AN - SCOPUS:85044751853
SN - 1862-8346
VL - 12
JO - Proteomics - Clinical Applications
JF - Proteomics - Clinical Applications
IS - 5
M1 - 1700012
ER -