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Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone
The adenosine 5′-triphosphate binding cassette subfamily B member (ABCB)11 gene is involved in bile transport, and mutations in this gene are associated with cholestasis and cholelithiasis. Therefore, the aim of the present study was to investigate the association between ABCB11 gene mutation and pr...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
D.A. Spandidos
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6297787/ https://www.ncbi.nlm.nih.gov/pubmed/30431138 http://dx.doi.org/10.3892/mmr.2018.9661 |
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author | Gan, Lang Pan, Shuguang Cui, Jinchi Bai, Jie Jiang, Peng He, Yu |
author_facet | Gan, Lang Pan, Shuguang Cui, Jinchi Bai, Jie Jiang, Peng He, Yu |
author_sort | Gan, Lang |
collection | PubMed |
description | The adenosine 5′-triphosphate binding cassette subfamily B member (ABCB)11 gene is involved in bile transport, and mutations in this gene are associated with cholestasis and cholelithiasis. Therefore, the aim of the present study was to investigate the association between ABCB11 gene mutation and primary intrahepatic stone (PIS)s and to investigate the mechanism through which ABCB11 gene mutations affect the expression of the corresponding protein. Mutations of the ABCB11 gene in 443 PIS patients and 560 healthy participants were detected by exon sequencing. The expression levels of ABCB11 mRNA and bile salt export pump (BSEP) protein in the liver tissues of patients with PISs were measured by quantitative polymerase chain reaction and western blot analysis. The mutant plasmids constructed by site-directed mutagenesis of the human BSEP gene were transfected into human embryonic kidney 293 (293) cells and Madin-Darby canine kidney (MDCK) cells, and the expression and distribution of rs118109635 of BSEP was measured. There were two significant mutations in the ABCB11 gene of the PIS patients compared with the healthy population; a missense mutation, rs118109635 (P=0.025), and a synonymous mutation, rs497692 (P=0.006). The two mutations were associated with the occurrence of preoperative jaundice (P=0.026, and P=0.011, respectively). The expression levels of BSEP in PIS patients with the missense mutation rs118109635 was decreased, whereas its mRNA expression levels remained unchanged. In PIS patients with the synonymous mutation rs497692, the expression levels of ABCB11 were decreased at both the mRNA and protein level. It was also found that mutation A865V reduced the expression levels of BSEP in 293 cells at the cellular level; its distribution in MDCK cell membranes was decreased, whereas its mRNA levels remained unchanged. The mutated loci at rs118109635 and rs497692 of the ABCB11 gene were correlated with PISs, causing a decreased expression of BSEP and reduced distribution of the protein in the cell membrane. Therefore, mutations at rs118109635 and rs497692 of the ABCB11 gene may be risk factors for PISs. |
format | Online Article Text |
id | pubmed-6297787 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | D.A. Spandidos |
record_format | MEDLINE/PubMed |
spelling | pubmed-62977872018-12-26 Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone Gan, Lang Pan, Shuguang Cui, Jinchi Bai, Jie Jiang, Peng He, Yu Mol Med Rep Articles The adenosine 5′-triphosphate binding cassette subfamily B member (ABCB)11 gene is involved in bile transport, and mutations in this gene are associated with cholestasis and cholelithiasis. Therefore, the aim of the present study was to investigate the association between ABCB11 gene mutation and primary intrahepatic stone (PIS)s and to investigate the mechanism through which ABCB11 gene mutations affect the expression of the corresponding protein. Mutations of the ABCB11 gene in 443 PIS patients and 560 healthy participants were detected by exon sequencing. The expression levels of ABCB11 mRNA and bile salt export pump (BSEP) protein in the liver tissues of patients with PISs were measured by quantitative polymerase chain reaction and western blot analysis. The mutant plasmids constructed by site-directed mutagenesis of the human BSEP gene were transfected into human embryonic kidney 293 (293) cells and Madin-Darby canine kidney (MDCK) cells, and the expression and distribution of rs118109635 of BSEP was measured. There were two significant mutations in the ABCB11 gene of the PIS patients compared with the healthy population; a missense mutation, rs118109635 (P=0.025), and a synonymous mutation, rs497692 (P=0.006). The two mutations were associated with the occurrence of preoperative jaundice (P=0.026, and P=0.011, respectively). The expression levels of BSEP in PIS patients with the missense mutation rs118109635 was decreased, whereas its mRNA expression levels remained unchanged. In PIS patients with the synonymous mutation rs497692, the expression levels of ABCB11 were decreased at both the mRNA and protein level. It was also found that mutation A865V reduced the expression levels of BSEP in 293 cells at the cellular level; its distribution in MDCK cell membranes was decreased, whereas its mRNA levels remained unchanged. The mutated loci at rs118109635 and rs497692 of the ABCB11 gene were correlated with PISs, causing a decreased expression of BSEP and reduced distribution of the protein in the cell membrane. Therefore, mutations at rs118109635 and rs497692 of the ABCB11 gene may be risk factors for PISs. D.A. Spandidos 2019-01 2018-11-15 /pmc/articles/PMC6297787/ /pubmed/30431138 http://dx.doi.org/10.3892/mmr.2018.9661 Text en Copyright: © Gan 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 Gan, Lang Pan, Shuguang Cui, Jinchi Bai, Jie Jiang, Peng He, Yu Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone |
title | Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone |
title_full | Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone |
title_fullStr | Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone |
title_full_unstemmed | Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone |
title_short | Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone |
title_sort | functional analysis of the correlation between abcb11 gene mutation and primary intrahepatic stone |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6297787/ https://www.ncbi.nlm.nih.gov/pubmed/30431138 http://dx.doi.org/10.3892/mmr.2018.9661 |
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