转录因子RPN4对白念珠菌药物敏感性的研究

中国真菌学杂志　 2015, Vol. 10 　Issue (2): 75-79.

转录因子RPN4对白念珠菌药物敏感性的研究

许洪涛^1,2, 唐若愚³, 屈莺歌^1,2, 王晓娟^1,2, 姜远英², 曹永兵², 颜天华¹

1. 中国药科大学药学院, 南京 211198;
2. 第二军医大学药学院新药研究中心, 上海 200433;
3. 上海海洋大学食品学院, 上海 201306

收稿日期:2015-03-13 出版日期:2015-04-28 发布日期:2015-04-28
通讯作者: 颜天华,E-mail:18915991229@126.com E-mail:18915991229@126.com
作者简介:许洪涛,男(汉族),硕士研究生在读.E-mail:xhthfut@163.com
基金资助:
国家自然科学基金 (81173100),上海市基础研究重点课题 (11JC1415400)

Analysis of relationship between transcription factor RPN4 and drug susceptibility of Candida albicans

XU Hong-tao^1,2, TANG Ruo-yu³, QU Ying-ge^1,2, WANG Xiao-juan^1,2, JIANG Yuan-ying², CAO Yong-bing², YAN Tian-hua¹

1. College of Pharmacy, China Pharmaceutical University, Nanjing 211198;
2. New Drug Research and Development Center, College of Pharmacy, The Second Military Medical University, Shanghai 200433;
3. College of Food Science and Technology, ShangHai Ocean University, Shanghai 201306

Received:2015-03-13 Online:2015-04-28 Published:2015-04-28

摘要/Abstract

摘要：

目的利用基因缺失菌库研究转录因子敲除后对白念珠菌药物敏感性的影响,并利用药物敏感性差异菌株初步考察可能的耐药性调控机制。方法微量液基稀释法测定最低抑菌浓度 (minimal inhibitory concentration,MIC);点板法 (spot assay)和生长曲线法验证菌株对氟康唑的敏感性;实时定量PCR (RT-PCR)法检测药物敏感性差异菌株中多药耐药基因CDR1和MDR1的表达,并通过罗丹明6G外排实验测定外排能力。结果亲本菌SN250对氟康唑表现为耐药,多药耐药基因CDR1和MDR1高表达,MIC₈₀大于16 μg/mL,大部分转录因子缺失菌与亲本菌SN250表现出相同的耐药性,但转录因子RPN4缺失菌株对氟康唑的敏感性升高,MIC₈₀降为0.5 μg/mL;多药耐药基因CDR1和MDR1的表达均降低,20 min和40 min时对罗丹明6G的外排能力降低。结论转录因子RPN4有可能通过促进多药耐药基因CDR1和MDR1表达和菌株外排能力而降低对药物的敏感性,但相关机制有待进一步深入研究。

关键词: 白念珠菌, 转录因子, 药物敏感性, 多药耐药基因

Abstract:

Objective To study the influence on the drug susceptibility of Candida albicans after transcription factor RPN4 was knocked out and the mechanism how this transcription factor regulates the drug susceptibility in Candida albicans.Methods The broth microdilution method was used to determine the minimal inhibitory concentrations (MICs) of 30 strains transcription factor knocked out Candida albicans .The drug susceptibility was reassured by spot assay and growth curve.The total RNA of parental strain SN250 and susceptible strains were extracted and the expression of multidrug resistance genes CDR1 and MDR1 was determined by real-time PCR,and the pump ability was examined by the ability of pump out rhodamine 6G.Results Compaired to the parental strain SN250,transcription factor RPN4 knocked out strain was more susceptible to fluconazole.The expression of CDR1 and MDR1 was lower than SN250 and the pump ability was also weaker than SN250.Conclusion The knockout of transcription factor PRN4 results susceptible to fluconazole in Candida albicans,and this transcription factor may be involved in the regulation of drug susceptibility in Candida albicans.

Key words: Candida albicans, transcription factors, drug susceptibility, multidrug resistance genes

中图分类号:

R379.4

许洪涛, 唐若愚, 屈莺歌, 王晓娟, 姜远英, 曹永兵, 颜天华. 转录因子RPN4对白念珠菌药物敏感性的研究[J]. 中国真菌学杂志, 2015, 10(2): 75-79.

XU Hong-tao, TANG Ruo-yu, QU Ying-ge, WANG Xiao-juan, JIANG Yuan-ying, CAO Yong-bing, YAN Tian-hua. Analysis of relationship between transcription factor RPN4 and drug susceptibility of Candida albicans[J]. Chinese Journal of Mycology, 2015, 10(2): 75-79.

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