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Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells

The inhibition of the activation of hepatic stellate cells (HSCs) and the induction of their apoptosis have been investigated as potential strategies to counteract the development and progression of liver fibrosis. Previous research has suggested that apoptosis-related protein in the transforming gr...

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Autores principales: Xu, Feifan, Han, Yuanlong, Zhu, Dandan, Tian, Hua, Zhu, Huiming, Ren, Jingjing, Gu, Delin, Duan, Yinong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5647026/
https://www.ncbi.nlm.nih.gov/pubmed/28791356
http://dx.doi.org/10.3892/mmr.2017.7172
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author Xu, Feifan
Han, Yuanlong
Zhu, Dandan
Tian, Hua
Zhu, Huiming
Ren, Jingjing
Gu, Delin
Duan, Yinong
author_facet Xu, Feifan
Han, Yuanlong
Zhu, Dandan
Tian, Hua
Zhu, Huiming
Ren, Jingjing
Gu, Delin
Duan, Yinong
author_sort Xu, Feifan
collection PubMed
description The inhibition of the activation of hepatic stellate cells (HSCs) and the induction of their apoptosis have been investigated as potential strategies to counteract the development and progression of liver fibrosis. Previous research has suggested that apoptosis-related protein in the transforming growth factor-β signaling pathway (ARTS) may serve a significant role in numerous cell types; however, little is known regarding its roles in HSCs. Total RNA was extracted from LX-2 cells, and the human full-length ARTS gene was obtained by reverse transcription-polymerase chain reaction and inserted into the pIRES2-EGFP cloning vector. Subsequently, the recombinant pIRES2-EGFP-ARTS plasmid was transfected into LX-2 cells by FuGENE 6 transfection reagent, and the expression of ARTS was detected by western blotting and fluorescent microscopy. In addition, the effects of pIRES2-EGFP-ARTS on the activation, apoptosis, viability and migration of LX-2 cells were assessed by western blot analysis, TUNEL staining, an MTT assay, and scratch and Transwell assays, respectively. The present results demonstrated that the pIRES2-EGFP-ARTS vector expressing human ARTS was successfully constructed, and the overexpression of ARTS contributed to enhance the apoptosis and inhibit the activation of human LX-2 HSCs. The present findings suggested that ARTS overexpression may have potential as a novel therapeutic strategy to reverse hepatic fibrosis.
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spelling pubmed-56470262017-10-24 Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells Xu, Feifan Han, Yuanlong Zhu, Dandan Tian, Hua Zhu, Huiming Ren, Jingjing Gu, Delin Duan, Yinong Mol Med Rep Articles The inhibition of the activation of hepatic stellate cells (HSCs) and the induction of their apoptosis have been investigated as potential strategies to counteract the development and progression of liver fibrosis. Previous research has suggested that apoptosis-related protein in the transforming growth factor-β signaling pathway (ARTS) may serve a significant role in numerous cell types; however, little is known regarding its roles in HSCs. Total RNA was extracted from LX-2 cells, and the human full-length ARTS gene was obtained by reverse transcription-polymerase chain reaction and inserted into the pIRES2-EGFP cloning vector. Subsequently, the recombinant pIRES2-EGFP-ARTS plasmid was transfected into LX-2 cells by FuGENE 6 transfection reagent, and the expression of ARTS was detected by western blotting and fluorescent microscopy. In addition, the effects of pIRES2-EGFP-ARTS on the activation, apoptosis, viability and migration of LX-2 cells were assessed by western blot analysis, TUNEL staining, an MTT assay, and scratch and Transwell assays, respectively. The present results demonstrated that the pIRES2-EGFP-ARTS vector expressing human ARTS was successfully constructed, and the overexpression of ARTS contributed to enhance the apoptosis and inhibit the activation of human LX-2 HSCs. The present findings suggested that ARTS overexpression may have potential as a novel therapeutic strategy to reverse hepatic fibrosis. D.A. Spandidos 2017-10 2017-08-04 /pmc/articles/PMC5647026/ /pubmed/28791356 http://dx.doi.org/10.3892/mmr.2017.7172 Text en Copyright: © Xu 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
Xu, Feifan
Han, Yuanlong
Zhu, Dandan
Tian, Hua
Zhu, Huiming
Ren, Jingjing
Gu, Delin
Duan, Yinong
Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells
title Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells
title_full Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells
title_fullStr Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells
title_full_unstemmed Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells
title_short Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells
title_sort construction of a recombinant pires2-egfp-arts plasmid and its effect on lx-2 cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5647026/
https://www.ncbi.nlm.nih.gov/pubmed/28791356
http://dx.doi.org/10.3892/mmr.2017.7172
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