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Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells

Rab5a, a key member of the Rab family of GTPases, was determined to be a regulator of vascular smooth muscle cell (VSMC) proliferation and migration. However, the exact regulatory mechanism remains unclear. As Rab5a has been shown to be associated with autophagy, which is essential for the conversio...

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Autores principales: Tan, Jin-Yun, Jia, Luo-Qi, Shi, Wei-Hao, He, Qing, Zhu, Lei, Yu, Bo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5101961/
https://www.ncbi.nlm.nih.gov/pubmed/27666726
http://dx.doi.org/10.3892/mmr.2016.5774
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author Tan, Jin-Yun
Jia, Luo-Qi
Shi, Wei-Hao
He, Qing
Zhu, Lei
Yu, Bo
author_facet Tan, Jin-Yun
Jia, Luo-Qi
Shi, Wei-Hao
He, Qing
Zhu, Lei
Yu, Bo
author_sort Tan, Jin-Yun
collection PubMed
description Rab5a, a key member of the Rab family of GTPases, was determined to be a regulator of vascular smooth muscle cell (VSMC) proliferation and migration. However, the exact regulatory mechanism remains unclear. As Rab5a has been shown to be associated with autophagy, which is essential for the conversion of VSMCs from a contractile to a synthetic phenotype in order to prevent cell death due to oxidative stress. The present study hypothesized that autophagy may be responsible for the proliferation and migration of VSMCs via the Rab5a protein. The aim of the present study was to evaluate the effect of Rab5a on autophagy in VSMCs. The human aorta vascular smooth muscle cell line, T/G HA-VSMCs, was treated with small interfering (si)RNA against Rab5a and/or platelet-derived growth factor (PDGF). Following treatment, the phenotype transition of the VSMCs was evaluated by detecting the mRNA and protien expression levels of VSMC molecular markers using reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. In addition, autophagy in VSMCs was evaluated by western blotting for autophagy-associated proteins, flow cytometry of acidic vesicular organelles, punctate fluorescence of microtubule associated protein light chain 3 and transmission electron microscopy of typical scattered double-membrane vacuolar structures. Additionally, the proliferation, migration, cell cycle and apoptotic response of VSMCs were detected by sulforhodamine B assay, transwell assay and flow cytometry, respectively. The results revealed that transfection with siRNA against Rab5a led to a significant decrease in Rab5a protein expression, while the reduced expression trend of Rab5a was rescued by intervention with PDGF. Furthermore, cells transfected with siRNA against Rab5a inhibited the autophagy of VSMCs. Downregulated Rab5a inhibited the phenotype transition of VSMCs. Additionally, downregulated Rab5a led to slowed cell growth, decreased numbers of migrated cells, decreased numbers of cells at the G0-G1 phase and a higher apoptosis rate. However, PDGF significantly rescued these phenomena caused by siRNA against Rab5a. These results indicated that Rab5a-mediated autophagy may regulate the phenotype transition and cell behavior of VSMCs through the activation of the extracellular-regulated kinase 1/2 signaling pathway.
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spelling pubmed-51019612016-11-22 Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells Tan, Jin-Yun Jia, Luo-Qi Shi, Wei-Hao He, Qing Zhu, Lei Yu, Bo Mol Med Rep Articles Rab5a, a key member of the Rab family of GTPases, was determined to be a regulator of vascular smooth muscle cell (VSMC) proliferation and migration. However, the exact regulatory mechanism remains unclear. As Rab5a has been shown to be associated with autophagy, which is essential for the conversion of VSMCs from a contractile to a synthetic phenotype in order to prevent cell death due to oxidative stress. The present study hypothesized that autophagy may be responsible for the proliferation and migration of VSMCs via the Rab5a protein. The aim of the present study was to evaluate the effect of Rab5a on autophagy in VSMCs. The human aorta vascular smooth muscle cell line, T/G HA-VSMCs, was treated with small interfering (si)RNA against Rab5a and/or platelet-derived growth factor (PDGF). Following treatment, the phenotype transition of the VSMCs was evaluated by detecting the mRNA and protien expression levels of VSMC molecular markers using reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. In addition, autophagy in VSMCs was evaluated by western blotting for autophagy-associated proteins, flow cytometry of acidic vesicular organelles, punctate fluorescence of microtubule associated protein light chain 3 and transmission electron microscopy of typical scattered double-membrane vacuolar structures. Additionally, the proliferation, migration, cell cycle and apoptotic response of VSMCs were detected by sulforhodamine B assay, transwell assay and flow cytometry, respectively. The results revealed that transfection with siRNA against Rab5a led to a significant decrease in Rab5a protein expression, while the reduced expression trend of Rab5a was rescued by intervention with PDGF. Furthermore, cells transfected with siRNA against Rab5a inhibited the autophagy of VSMCs. Downregulated Rab5a inhibited the phenotype transition of VSMCs. Additionally, downregulated Rab5a led to slowed cell growth, decreased numbers of migrated cells, decreased numbers of cells at the G0-G1 phase and a higher apoptosis rate. However, PDGF significantly rescued these phenomena caused by siRNA against Rab5a. These results indicated that Rab5a-mediated autophagy may regulate the phenotype transition and cell behavior of VSMCs through the activation of the extracellular-regulated kinase 1/2 signaling pathway. D.A. Spandidos 2016-11 2016-09-26 /pmc/articles/PMC5101961/ /pubmed/27666726 http://dx.doi.org/10.3892/mmr.2016.5774 Text en Copyright: © Tan et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Tan, Jin-Yun
Jia, Luo-Qi
Shi, Wei-Hao
He, Qing
Zhu, Lei
Yu, Bo
Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells
title Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells
title_full Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells
title_fullStr Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells
title_full_unstemmed Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells
title_short Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells
title_sort rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5101961/
https://www.ncbi.nlm.nih.gov/pubmed/27666726
http://dx.doi.org/10.3892/mmr.2016.5774
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