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Location of NLS-RARα protein in NB4 cell and nude mice
In the majority of acute promyelocytic leukemia (APL) cases, translocons produce a promyelocytic leukemia protein-retinoic acid receptor α (PML-RARα) fusion gene. Studies have reported that neutrophil elastase (NE) cleaves bcr-1-derived PML-RAα in early myeloid cells, leaving only the nuclear locali...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
D.A. Spandidos
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5403253/ https://www.ncbi.nlm.nih.gov/pubmed/28454360 http://dx.doi.org/10.3892/ol.2017.5706 |
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author | Wang, Hui Yang, Rong Zhong, Liang Zhu, Xin-Yu Ma, Peng-Peng Yang, Xiao-Qun Jiang, Kai-Ling Liu, Bei-Zhong |
author_facet | Wang, Hui Yang, Rong Zhong, Liang Zhu, Xin-Yu Ma, Peng-Peng Yang, Xiao-Qun Jiang, Kai-Ling Liu, Bei-Zhong |
author_sort | Wang, Hui |
collection | PubMed |
description | In the majority of acute promyelocytic leukemia (APL) cases, translocons produce a promyelocytic leukemia protein-retinoic acid receptor α (PML-RARα) fusion gene. Studies have reported that neutrophil elastase (NE) cleaves bcr-1-derived PML-RAα in early myeloid cells, leaving only the nuclear localization signal (NLS) of PML attached to RARα. NLS-RARα promotes cell growth and inhibits differentiation in response to ATRA. However, the mechanisms by which NLS-RARα affects cell biological characteristics are yet to be fully elucidated. The present study found that the location of RARαwas altered after it was cleaved by NE. Firstly, NE was overexpressed during the preparation of recombinant plasmid NB-4/pCMV6-NE-Myc to cleave PML-RARα. The total protein expression levels of myc and NE and expression levels of NLS-RARα in nucleoprotein were detected by western blotting. Location of NLS-RARα protein was detected by immunofluorescence and confocal laser scanning. Secondly, a nude mice model was constructed and NE protein, NLS-RARα and RARα protein assays, and the location of NLS-RARα and RARα proteins were assessed as described. The present results showed that, compared with the control groups, the location of NLS-RARα protein was predominantly detected in the nucleus, whereas RARα was mainly distributed in the cytoplasm. These findings were consistent with those of the nude mice model, and these may be used as a foundation to explain the occurrence mechanism of APL. |
format | Online Article Text |
id | pubmed-5403253 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | D.A. Spandidos |
record_format | MEDLINE/PubMed |
spelling | pubmed-54032532017-04-27 Location of NLS-RARα protein in NB4 cell and nude mice Wang, Hui Yang, Rong Zhong, Liang Zhu, Xin-Yu Ma, Peng-Peng Yang, Xiao-Qun Jiang, Kai-Ling Liu, Bei-Zhong Oncol Lett Articles In the majority of acute promyelocytic leukemia (APL) cases, translocons produce a promyelocytic leukemia protein-retinoic acid receptor α (PML-RARα) fusion gene. Studies have reported that neutrophil elastase (NE) cleaves bcr-1-derived PML-RAα in early myeloid cells, leaving only the nuclear localization signal (NLS) of PML attached to RARα. NLS-RARα promotes cell growth and inhibits differentiation in response to ATRA. However, the mechanisms by which NLS-RARα affects cell biological characteristics are yet to be fully elucidated. The present study found that the location of RARαwas altered after it was cleaved by NE. Firstly, NE was overexpressed during the preparation of recombinant plasmid NB-4/pCMV6-NE-Myc to cleave PML-RARα. The total protein expression levels of myc and NE and expression levels of NLS-RARα in nucleoprotein were detected by western blotting. Location of NLS-RARα protein was detected by immunofluorescence and confocal laser scanning. Secondly, a nude mice model was constructed and NE protein, NLS-RARα and RARα protein assays, and the location of NLS-RARα and RARα proteins were assessed as described. The present results showed that, compared with the control groups, the location of NLS-RARα protein was predominantly detected in the nucleus, whereas RARα was mainly distributed in the cytoplasm. These findings were consistent with those of the nude mice model, and these may be used as a foundation to explain the occurrence mechanism of APL. D.A. Spandidos 2017-04 2017-02-08 /pmc/articles/PMC5403253/ /pubmed/28454360 http://dx.doi.org/10.3892/ol.2017.5706 Text en Copyright: © Wang 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 Wang, Hui Yang, Rong Zhong, Liang Zhu, Xin-Yu Ma, Peng-Peng Yang, Xiao-Qun Jiang, Kai-Ling Liu, Bei-Zhong Location of NLS-RARα protein in NB4 cell and nude mice |
title | Location of NLS-RARα protein in NB4 cell and nude mice |
title_full | Location of NLS-RARα protein in NB4 cell and nude mice |
title_fullStr | Location of NLS-RARα protein in NB4 cell and nude mice |
title_full_unstemmed | Location of NLS-RARα protein in NB4 cell and nude mice |
title_short | Location of NLS-RARα protein in NB4 cell and nude mice |
title_sort | location of nls-rarα protein in nb4 cell and nude mice |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5403253/ https://www.ncbi.nlm.nih.gov/pubmed/28454360 http://dx.doi.org/10.3892/ol.2017.5706 |
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