耐氟康唑光滑念珠菌耐药机制研究

Research on the mechanisms of fluconazole resistance in clinical isolates of Candida glabrata

目的探讨ERG11、CDR1和CDR2基因差异表达在耐氟康唑光滑念珠菌耐药性形成中的作用。方法采用实时荧光定量逆转录聚合酶链反应(PCR)对临床分离的耐氟康唑药物株、剂量依赖性敏感株和敏感株共22株光滑念珠菌的ERG11、CDR1和CDR2基因表达的mRNA进行相对定量,以2-△△Ct表示试验组目的基因的表达相对于对照组的变化倍数。所有数据采用R(2.15.2)软件进行统计学分析。结果耐药株及剂量依赖性敏感株CDR1、CDR2以及ERG11基因的mRNA表达量均高于敏感株(P<0.05),且随着对氟康唑耐药程度增加而增加。结论 CDR1、CDR2及ERG11基因表达上调是光滑念珠菌临床分离株对氟康唑耐药的主要分子机制。

Objective To investigate the role of the expressions of ERG11, CDR1 and CDR2 genes in the fluconazole resistance to Candida glabrata. Methods Real-time fluorescence quantitation reverse transcription polymerase chain reaction （PCR） was used to evaluate the mRNA expressions of ERGll, CDR1 and CDR2 genes in 22 clinical isolates including fluconazole-resistant, dose-dependent-sensitive and fluconazole-sensitive Candida glabrata.2 -△△ct was used to determine the relative multiples of the changes between the experimental group and the control group. Statistical analysis was performed with R （2.15.2） software. Results CDR1, CDR2 and ERGll gene mRNA expressions were significantly higher in the fluconazole-resistant isolates and dose-dependent-sensitive isolates than in the fluconazole-sensitive isolates （ P 〈 0.05 ）. With the decreasing of susceptibilities to flueonazole, the expressions of CDR1, CDR2 and ERG11 genes appeared to increase. Conduslons The upregulation of CDR1, CDR2 and ERG11 genes is a major molecular mechanism of flueonazole resistance in clinical isolates of Candida glabrata.