Screening of fluconazole resistance-related genes in <italic style="font-style: italic">Trichosporon asahii</italic> with induced fluconazole resistance <italic style="font-style: italic">in vitro</italic>

Zhikuan XIA; Jing GUO; Dequan ZHANG; Yong LU; Junhong AO; Rongya YANG

doi:10.16016/j.1000-5404.202001156

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Screening of fluconazole resistance-related genes in Trichosporon asahii with induced fluconazole resistance in vitro

Basic Medicine | 更新时间：2026-01-09

- Screening of fluconazole resistance-related genes in Trichosporon asahii with induced fluconazole resistance in vitro
- Journal of Third Military Medical University Vol. 42, Issue 10, Pages: 996-1001(2020)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)研究生院
  2.100700 北京，解放军总医院第七医学中心全军皮肤损伤修复研究所
  3.100120 北京，军委政治工作部原机关门诊部
  4.150001 哈尔滨，93199 部队门诊部
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81271764);the General Program of National Natural Science Foundation of China(81571972);the General Program of Natural Science Foundation of Beijing(7202201)
- DOI：10.16016/j.1000-5404.202001156
  CLC： R379.9;R394-33;R915
- Received：07 January 2020，
  
  Revised：2020-03-05，
  
  Published：30 May 2020
- 稿件说明：
移动端阅览
Zhikuan XIA, Jing GUO, Dequan ZHANG, et al. Screening of fluconazole resistance-related genes in Trichosporon asahii with induced fluconazole resistance in vitro[J]. Journal of Third Military Medical University, 2020, 42(10): 996-1001.
DOI：

Zhikuan XIA, Jing GUO, Dequan ZHANG, et al. Screening of fluconazole resistance-related genes in Trichosporon asahii with induced fluconazole resistance in vitro[J]. Journal of Third Military Medical University, 2020, 42(10): 996-1001. DOI： 10.16016/j.1000-5404.202001156.

摘要

目的

筛选阿萨希毛孢子菌氟康唑耐药相关基因，为研究该菌氟康唑耐药机制及开发新型抗真菌剂提供理论依据。

方法

将临床分离的氟康唑敏感阿萨希毛孢子菌通过不断倍增氟康唑药物浓度的方法进行体外诱导培养，直至药物浓度≥128 μg/mL，再进行无药培养30代，测定其MIC值。分别提取敏感菌株和诱导耐药菌株的总RNA，构建RNA文库，转录组测序后进行生物信息学分析，寻找差异表达基因。相同方法诱导另外2株敏感菌株为耐药菌株，并对3对敏感与耐药菌株的基因组进行测序，比较ERG11基因序列。

结果

成功获得稳定的诱导耐药菌株，转录组发现耐药菌株中与药物转运相关基因、麦角固醇合成相关基因、抗氧化基因等上调表达，其中氟康唑靶酶编码基因ERG11基因表达上调了3.7倍。而基因组测序证实3株耐药菌株中ERG11基因的碱基序列未发生突变。

结论

阿萨希毛孢子菌耐药分子机制复杂，麦角固醇合成途径中的基因，特别是ERG11基因上调表达是其主要机制之一，同时还与外排转运基因、抗氧化基因等有关。

Abstract

Objective

To screen the genes associated with fluconazole resistance in

Trichosporon asahii

to facilitate mechanistic studies of fluconazole resistance and the development of new antifungal agents.

Methods

Three clinical isolates of fluconazole-sensitive

Trichosporon asahii

were cultured in the presence of increasing concentrations of fluconazole until the drug concentration reached 128 μg/mL. After further culture for 30 generations in the absence of fluconazole

the MIC value of the induced bacteria was measured. The total RNA was extracted from both the sensitive and resistant strains of the bacteria for construction of the stranded RNA-Seq libraries and transcriptome sequencing to identify the differentially expressed genes (DEGs) in the resistant strains. The genomes of the 3 sensitive and resistant strains were sequenced for comparison of the sequence of the differentially expressed ERG11 gene.

Results

We successfully obtained stable

Trichosporon asahii

strains with fluconazole resistance. Transcriptome sequencing data showed that the genes associated with drug transport

ergosterol synthesis and antioxidation were up-regulated in t

he resistant strains

and among them ERG11 gene

the target gene of fluconazole

was up-regulated by 3.7 folds. Genomic sequencing proved no mutations in the sequence of ERG11 gene in the 3 resistant strains.

Conclusion

The molecular mechanism for drug resistance in

Trichosporon asahii

is complex. The upregulation of ergosterol synthesis pathway

especially ERG11 gene

along with other genes associated with efflux transfer and oxidative stress

is one of the main mechanisms for drug resistance in

Trichosporon asahii

关键词

Keywords

references

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Screening of fluconazole resistance-related genes in Trichosporon asahii with induced fluconazole resistance in vitro

DOI：10.16016/j.1000-5404.202001156

摘要

Abstract

关键词

Keywords

references