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Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei

OBJECTIVE: To study the main molecular mechanisms responsible for itraconazole resistance in clinical isolates of Candida krusei. METHODS: The 14α-demethylases encoded by ERG11 gene in the 16 C.krusei clinical isolates were amplified by polymerase chain reaction (PCR), and their nucleotide sequences...

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Autores principales: He, Xiaoyuan, Zhao, Mingfeng, Chen, Jinyan, Wu, Rimao, Zhang, Jianlei, Cui, Rui, Jiang, Yanyu, Chen, Jie, Cao, Xiaoli, Xing, Yi, Zhang, Yuchen, Meng, Juanxia, Deng, Qi, Sui, Tao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4550294/
https://www.ncbi.nlm.nih.gov/pubmed/26308936
http://dx.doi.org/10.1371/journal.pone.0136185
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author He, Xiaoyuan
Zhao, Mingfeng
Chen, Jinyan
Wu, Rimao
Zhang, Jianlei
Cui, Rui
Jiang, Yanyu
Chen, Jie
Cao, Xiaoli
Xing, Yi
Zhang, Yuchen
Meng, Juanxia
Deng, Qi
Sui, Tao
author_facet He, Xiaoyuan
Zhao, Mingfeng
Chen, Jinyan
Wu, Rimao
Zhang, Jianlei
Cui, Rui
Jiang, Yanyu
Chen, Jie
Cao, Xiaoli
Xing, Yi
Zhang, Yuchen
Meng, Juanxia
Deng, Qi
Sui, Tao
author_sort He, Xiaoyuan
collection PubMed
description OBJECTIVE: To study the main molecular mechanisms responsible for itraconazole resistance in clinical isolates of Candida krusei. METHODS: The 14α-demethylases encoded by ERG11 gene in the 16 C.krusei clinical isolates were amplified by polymerase chain reaction (PCR), and their nucleotide sequences were determined to detect point mutations. Meanwhile, ERG11 and efflux transporters (ABC1 and ABC2) genes were determined by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) for their expression in itraconazole-resistant (R), itraconazole-susceptible dose dependent (SDD) and itraconazole-susceptible (S) C.krusei at the mRNA level. RESULTS: We found 7-point mutations in ERG11 gene of all the C.krusei clinical isolates, including 6 synonymous mutations and 1 missense mutation (C44T). However, the missense mutation was found in the three groups. The mRNA levels of ERG11 gene in itraconazole-resistant isolates showed higher expression compared with itraconazole-susceptible dose dependent and itraconazole-susceptible ones (P = 0.015 and P = 0.002 respectively). ABC2 gene mRNA levels in itraconazole-resistant group was significantly higher than the other two groups, and the levels of their expression in the isolates appeared to increase with the decrease of susceptibility to itraconazole (P = 0.007 in SDD compared with S, P = 0.016 in SDD with R, and P<0.001 in S with R respectively). While ABC1 gene presented lower expression in itraconazole resistant strains. However, the mRNA levels of ERG11, ABC1 and ABC2 in a C.krusei (CK10) resistant to both itraconazole and voriconazole were expressed highest in all the itraconazole-resistant isolates. CONCLUSIONS: There are ERG11 gene polymorphisms in clinical isolates of C.krusei. ERG11 gene mutations may not be involved in the development of itraconazole resistance in C.krusei. ERG11 and ABC2 overexpression might be responsible for the acquired itraconazole resistance of these clinical isolates.
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spelling pubmed-45502942015-09-01 Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei He, Xiaoyuan Zhao, Mingfeng Chen, Jinyan Wu, Rimao Zhang, Jianlei Cui, Rui Jiang, Yanyu Chen, Jie Cao, Xiaoli Xing, Yi Zhang, Yuchen Meng, Juanxia Deng, Qi Sui, Tao PLoS One Research Article OBJECTIVE: To study the main molecular mechanisms responsible for itraconazole resistance in clinical isolates of Candida krusei. METHODS: The 14α-demethylases encoded by ERG11 gene in the 16 C.krusei clinical isolates were amplified by polymerase chain reaction (PCR), and their nucleotide sequences were determined to detect point mutations. Meanwhile, ERG11 and efflux transporters (ABC1 and ABC2) genes were determined by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) for their expression in itraconazole-resistant (R), itraconazole-susceptible dose dependent (SDD) and itraconazole-susceptible (S) C.krusei at the mRNA level. RESULTS: We found 7-point mutations in ERG11 gene of all the C.krusei clinical isolates, including 6 synonymous mutations and 1 missense mutation (C44T). However, the missense mutation was found in the three groups. The mRNA levels of ERG11 gene in itraconazole-resistant isolates showed higher expression compared with itraconazole-susceptible dose dependent and itraconazole-susceptible ones (P = 0.015 and P = 0.002 respectively). ABC2 gene mRNA levels in itraconazole-resistant group was significantly higher than the other two groups, and the levels of their expression in the isolates appeared to increase with the decrease of susceptibility to itraconazole (P = 0.007 in SDD compared with S, P = 0.016 in SDD with R, and P<0.001 in S with R respectively). While ABC1 gene presented lower expression in itraconazole resistant strains. However, the mRNA levels of ERG11, ABC1 and ABC2 in a C.krusei (CK10) resistant to both itraconazole and voriconazole were expressed highest in all the itraconazole-resistant isolates. CONCLUSIONS: There are ERG11 gene polymorphisms in clinical isolates of C.krusei. ERG11 gene mutations may not be involved in the development of itraconazole resistance in C.krusei. ERG11 and ABC2 overexpression might be responsible for the acquired itraconazole resistance of these clinical isolates. Public Library of Science 2015-08-26 /pmc/articles/PMC4550294/ /pubmed/26308936 http://dx.doi.org/10.1371/journal.pone.0136185 Text en © 2015 He et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
He, Xiaoyuan
Zhao, Mingfeng
Chen, Jinyan
Wu, Rimao
Zhang, Jianlei
Cui, Rui
Jiang, Yanyu
Chen, Jie
Cao, Xiaoli
Xing, Yi
Zhang, Yuchen
Meng, Juanxia
Deng, Qi
Sui, Tao
Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei
title Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei
title_full Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei
title_fullStr Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei
title_full_unstemmed Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei
title_short Overexpression of Both ERG11 and ABC2 Genes Might Be Responsible for Itraconazole Resistance in Clinical Isolates of Candida krusei
title_sort overexpression of both erg11 and abc2 genes might be responsible for itraconazole resistance in clinical isolates of candida krusei
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4550294/
https://www.ncbi.nlm.nih.gov/pubmed/26308936
http://dx.doi.org/10.1371/journal.pone.0136185
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