LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion
Monocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adh...
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doaj-0955c5e7166348ecb4dbf4f30e0042e02020-11-24T20:58:44ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2018-07-01510.3389/fmolb.2018.00067327021LPS Induces mTORC1 and mTORC2 Activation During Monocyte AdhesionMarcelle C. Ribeiro0Diogo B. Peruchetti1Leandro S. Silva2João L. Silva-Filho3Mariana C. Souza4Maria das Graças Henriques5Celso Caruso-Neves6Celso Caruso-Neves7Ana Acacia S. Pinheiro8Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrazilInstituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrazilInstituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrazilInstituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrazilFundação Oswaldo Cruz, Instituto de Tecnologia em Fármacos, Rio de Janeiro, BrazilFundação Oswaldo Cruz, Instituto de Tecnologia em Fármacos, Rio de Janeiro, BrazilInstituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrazilInstituto Nacional de Ciência e Tecnologia em Medicina Regenerativa, Rio de Janeiro, BrazilInstituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrazilMonocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adhesion under static and flow conditions. We observed that 1.0 μg/mL LPS increased PI3K/mTORC2 pathway and PKC activity after 1 h of incubation. WYE-354 10−6 M (mTORC2/mTORC1 inhibitor) and 10−6 M wortmannin avoided monocyte adhesion in culture plates. In addition, WYE also blocked LPS-induced CD11a expression. Interestingly, rapamycin and WYE-354 blocked both LPS-induced monocyte adhesion in a cell monolayer and actin cytoskeleton rearrangement, confirming mTORC1 involvement in this process. Once activated, PKC activates mTORC1/S6K pathway in a similar effect observed to LPS. Activation of the mTORC1/S6K pathway was attenuated by 10−6 M U0126, an MEK/ERK inhibitor, and 10−6 M calphostin C, a PKC inhibitor, indicating that the MEK/ERK/TSC2 axis acts as a mediator. In agreement, 80 nM PMA (a PKC activator) mimicked the effect of LPS on the activation of the MEK/ERK/TSC2/mTORC1/S6K pathway, monocyte adhesion to ECV cells and actin cytoskeleton rearrangement. Our findings show that LPS induces activation of mTOR complexes. This signaling pathway led to integrin expression and cytoskeleton rearrangement resulting in monocyte adhesion. These results describe a new molecular mechanism involved in monocyte adhesion in immune-based diseases.https://www.frontiersin.org/article/10.3389/fmolb.2018.00067/fulllipopolysaccharidemTORC1mTORC2protein kinase Cphorbol esterhuman acute monocytic leukemia cell line |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Marcelle C. Ribeiro Diogo B. Peruchetti Leandro S. Silva João L. Silva-Filho Mariana C. Souza Maria das Graças Henriques Celso Caruso-Neves Celso Caruso-Neves Ana Acacia S. Pinheiro |
spellingShingle |
Marcelle C. Ribeiro Diogo B. Peruchetti Leandro S. Silva João L. Silva-Filho Mariana C. Souza Maria das Graças Henriques Celso Caruso-Neves Celso Caruso-Neves Ana Acacia S. Pinheiro LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion Frontiers in Molecular Biosciences lipopolysaccharide mTORC1 mTORC2 protein kinase C phorbol ester human acute monocytic leukemia cell line |
author_facet |
Marcelle C. Ribeiro Diogo B. Peruchetti Leandro S. Silva João L. Silva-Filho Mariana C. Souza Maria das Graças Henriques Celso Caruso-Neves Celso Caruso-Neves Ana Acacia S. Pinheiro |
author_sort |
Marcelle C. Ribeiro |
title |
LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion |
title_short |
LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion |
title_full |
LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion |
title_fullStr |
LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion |
title_full_unstemmed |
LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion |
title_sort |
lps induces mtorc1 and mtorc2 activation during monocyte adhesion |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Molecular Biosciences |
issn |
2296-889X |
publishDate |
2018-07-01 |
description |
Monocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adhesion under static and flow conditions. We observed that 1.0 μg/mL LPS increased PI3K/mTORC2 pathway and PKC activity after 1 h of incubation. WYE-354 10−6 M (mTORC2/mTORC1 inhibitor) and 10−6 M wortmannin avoided monocyte adhesion in culture plates. In addition, WYE also blocked LPS-induced CD11a expression. Interestingly, rapamycin and WYE-354 blocked both LPS-induced monocyte adhesion in a cell monolayer and actin cytoskeleton rearrangement, confirming mTORC1 involvement in this process. Once activated, PKC activates mTORC1/S6K pathway in a similar effect observed to LPS. Activation of the mTORC1/S6K pathway was attenuated by 10−6 M U0126, an MEK/ERK inhibitor, and 10−6 M calphostin C, a PKC inhibitor, indicating that the MEK/ERK/TSC2 axis acts as a mediator. In agreement, 80 nM PMA (a PKC activator) mimicked the effect of LPS on the activation of the MEK/ERK/TSC2/mTORC1/S6K pathway, monocyte adhesion to ECV cells and actin cytoskeleton rearrangement. Our findings show that LPS induces activation of mTOR complexes. This signaling pathway led to integrin expression and cytoskeleton rearrangement resulting in monocyte adhesion. These results describe a new molecular mechanism involved in monocyte adhesion in immune-based diseases. |
topic |
lipopolysaccharide mTORC1 mTORC2 protein kinase C phorbol ester human acute monocytic leukemia cell line |
url |
https://www.frontiersin.org/article/10.3389/fmolb.2018.00067/full |
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