荧光定量PCR检测不同状态下白念珠菌CPH1、EFG1基因的表达

中国真菌学杂志　 2012, Vol. 　Issue (2): 77-81.

荧光定量PCR检测不同状态下白念珠菌CPH1、EFG1基因的表达

阳隽¹, 张天托², 朱家馨²

1. 广州医学院第二附属医院呼吸内科, 广州 510260;
2. 中山大学第三附属医院呼吸内科, 广州 510630

收稿日期:2011-11-09 出版日期:2012-04-28 发布日期:2012-04-28
作者简介:阳隽,女(汉族),博士,讲师.E-mail:yangjun9909@126.com

Detecting CPH1,EFG1 gene expression of planktonic cells and biofilm of Candida albicans with fluorescent quantitative PCR assay

YANG Jun¹, ZHANG Tian-tuo², ZHU Jia-xin²

1. Department of Respiratory Medicine, The Second Affiliated Hospital of GuangZhou Medical University, Guangzhou 510260;
2. Department of Respiratory Medicine, The third affiliated Hospital of Sun Yat-sen University, Guangzhou 510630

Received:2011-11-09 Online:2012-04-28 Published:2012-04-28

摘要/Abstract

摘要： 目的检测转录因子CPH1和EFG1基因在游离态及生物膜态呼吸道白念珠菌临床分离株的表达差异,探讨其在生物膜形成过程中的作用。方法选取10 株白念珠菌临床分离株,分别提取游离态及生物膜态白念珠菌总RNA,用荧光定量PCR的方法测定两种状态下CPH1和EFG1基因的表达,用△△Ct的方法计算其相对表达量。结果白念珠菌生物膜态转录因子EFG1的表达是游离态表达水平的1.42~7.14倍,差异有显著意义(P<0.05),而转录因子CPH1的表达有8株菌生物膜态较游离态增高,1株降低,1株无明显变化,差异无显著意义(P>0.05)。结论白念珠临床株转录因子CPH1和EFG1参与生物膜形成的调控,并需在体内实验中进一步研究。

关键词: 白念珠菌, 生物膜, 转录因子

Abstract: Objective To investigate the role of gene expression of transcription factor CPH1 and EFG1 in biofilm formation by detecting the difference of CPH1,EFG1 mRNA expression between biofilm and planktonic cells of C.albicans.Methods Ten C.albicans strains isolated from respiratory tract were used. The total RNA of planktonic and biofilm cells were extracted separately, the mRNA expression of CPH1,EFG1 were measured with fluorescent quantitative PCR assay.The △△Ct that show relative value of mRNA expression were calculated.Results In this study,the mRNA expression of EFG1 in biofilm cells was 1.42~7.14 times higher than that in planktonic cells(P<0.05).The mRNA expression of CPH1 was up-regulated in biofilm cells of 8 isolates, down-regulated in 1 isolate and show no difference in 1 isolate(P>0.05).Conclusion The study show that transcription factors CPH1,EFG1 participated in the regulation of biofilm formation in clinical isolates, and further research in vivo is needed to provide more proffs.

Key words: Candida albicans, biofilm, transcription factor

中图分类号:

R379.4

阳隽, 张天托, 朱家馨. 荧光定量PCR检测不同状态下白念珠菌CPH1、EFG1基因的表达[J]. 中国真菌学杂志, 2012, (2): 77-81.

YANG Jun, ZHANG Tian-tuo, ZHU Jia-xin. Detecting CPH1,EFG1 gene expression of planktonic cells and biofilm of Candida albicans with fluorescent quantitative PCR assay[J]. Chinese Journal of Mycology, 2012, (2): 77-81.

参考文献

[1] Uppuluri P, Dinakaran H, Thomas DP, et al. Characteristics of Candida albicans biofilms grown in a synthetic urine medium[J]. J Clin Microbiol, 2009, 47(12): 4078-4083.
[2] Nailisl H, Kucharíková S,Ricicová M, et al. Real-time PCR expression profiling of genes encoding potential virulence factors in Candida albicans biofilms: identification of model-dependent and -independent gene expression[J]. BMC Microbiol,2010, 10(16):114-125.
[3] Nailis H, Coenye T, Van Nieuwerburgh F, et al. Development and evaluation of different normalization strategies for gene expression studies in Candida albicans biofilms by real-time PCR[J]. BMC Mol Biol,2006, 7(4):7-25.
[4] García-Sánchez S, Aubert S, Iraqui I, et al.Candida albicans biofilms: a developmental state associated with specific and stable gene expression patterns[J]. Eukaryot Cell,2004,3(2):536-545.
[5] Midkiff J, Borochoff-Porte N, White D, et al.Small molecule inhibitors of the Candida albicans budded-to-hyphal transition act through multiple signaling pathways[J]. PLoS One,2011,6(9):e25395.
[6] Nobile CJ,Mitchell AP.Genetics and genomics of Candida albicans biofilm formation[J].Cell Microbiol,2006,8(9):1382-1391.
[7] Kumamoto CA.A contact-activated kinase signals Candida albicans invasive growth and biofilm development[J].Proc Natl Acad Sci USA,2005,102(15):5576-5581.
[8] 陈曦,蒋云爱,孙小刚,等.MAPK和cAMP途径对白色念珠菌毒性与基因表达的影响 [J]. 科学通报, 2001,46(2):104-109.
[9] Dieterich C, SchandarM, NollM, et al.In vitro reconstructed human epithelia reveal contributions of Candida albicans EFG1 and CPH1 to adhesion and invasion[J]. Microbiology, 2002, 148(pt2):497-506.
[10] Lane S, Birse C, Zhou S, et al. DNA srray studies demonstrate convergent regulation of virulence factors by Cph1, Cph2, and Efg1 in Candida albicans[J]. J Biol Chem,2001,276(52):48988-48996.
[11] Stichternoth C, Ernst JF. Hypoxic adaptation by Efg1 regulates biofilm formation by Candida albicans[J].Appl Environ Microbiol,2009, 75(11):3663-3672.

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