UVA对白念珠菌生物膜作用的转录组学研究

中国真菌学杂志　 2017, Vol. 12 　Issue (6): 332-336.

UVA对白念珠菌生物膜作用的转录组学研究

李景¹, 朱丹婷², 俞立英¹

1. 复旦大学附属华山医院口腔科, 上海 200040;
2. 静安区牙病防治所, 上海 200040

收稿日期:2017-09-12 出版日期:2017-12-28 发布日期:2017-12-28
作者简介:李景,男(汉族),博士,主治医师.E-mail:lijing_joyo@163.com
基金资助:
上海市卫生计生委青年科研项目（20134y077）

Transcriptome analysis of Candida biofilm formation exposed to Ultraviolet A

LI Jing¹, ZHU Dan-ting², YU Li-ying¹

1. Department of Dentistry, Huashan Hospital, Fudan University, Shanghai 200040;
2. Shanghai Jing-an Dental Clinic, Shanghai 200040

Received:2017-09-12 Online:2017-12-28 Published:2017-12-28

摘要/Abstract

摘要：

目的应用转录组技术，对UVA照射后的白念珠菌菌丝及生物膜进行基因表达分析。方法体外构建白念珠菌（ATCC90028）生物膜，采用UVA-LED抗菌设备（复旦大学光电所制，波长365 nm，强度13 mW/cm²，频率100 Hz），照射白念珠菌生物膜5 min、10 min、15 min，对照组不光照。用转录组技术分析UVA实验组和对照组之间差异表达的基因，进行GO、KEGG遗传差异富集分析。结果 UVA光照组相对于对照组有大量差异表达基因，核糖体合成RIO2基因、氨基糖和核苷糖代谢相关基因MCR1、CBR1、内质网蛋白加工途径相关基因SHP1显著上调；RNA降解途径相关基因MPP6和细胞内吞作用相关基因CHMP6显著下调。结论 UVA直接或间接调节白念珠菌菌丝及生物膜相关生理代谢途径，参与诱发细胞凋亡过程。

关键词: 白念珠菌, UVA, RNA-seq测序, 差异表达基因

Abstract:

Objective To explore the genetic expression of hyphal growth and biofilm formation of Candida albicans exposed to ultraviolet A using the transcriptome analysis.Methods In vitro Candida albicans (ATCC90028) biofilms were irradiated by a new type of LED-UVA antimicrobial device (wavelength:365 nm,intensity:13 mW/cm²,frequency:100 Hz).The differentially expressed genes between the irradiated groups and the control group were detected by RNA-Seq technology analysis and GO,KEGG enrichment analysis of genetic differences.Results A total of 1221 gene expression levels exhibited a significant difference,including 862 up-regulated genes,359 down-regulated genes.The expressions of MCR1,CBR1,MUQ1 and SHP1 were significantly increased.The expressions of RIO2 and MPP were significantly decresed.Conclusion UVA could induce apoptosis of Candia albicans by un-regulating the expression of MCR1,CBR1,MUQ1 and SHP1 and inhibit hyphal growth and development by un-regulating the expression of RIO2 and MPP.

Key words: Candida albicans, UVA, RNA-seq sequencing, differentially expressed genes

中图分类号:

R379.4

李景, 朱丹婷, 俞立英. UVA对白念珠菌生物膜作用的转录组学研究[J]. 中国真菌学杂志, 2017, 12(6): 332-336.

LI Jing, ZHU Dan-ting, YU Li-ying. Transcriptome analysis of Candida biofilm formation exposed to Ultraviolet A[J]. Chinese Journal of Mycology, 2017, 12(6): 332-336.

参考文献

[1] Whiteway M,Bachewich C.Morphogenesis in Candida albicans[J].Annu Rev Microbiol,2007,61:529-553.
[2] 孙漩嵘.白念珠菌新基因IPF19998的氧化还原相关功能研究[D].上海:第二军医大学,药理学,2010.
[3] Ikner A,Shiozaki K.Yeast signaling pathways in the oxidative stress response[J].Mutat Res,2005,569(1-2):13-27.
[4] Li J,Hirota K,Yumoto H,et al.Enhanced germicidal effects of pulsed UV-LED irradiation on biofilms[J].J Appl Microbiol,2010,109(6):2183-2190.
[5] Amorim-Vaz S,Tran Vdu T,Pradervand S,et al.RNA enrichment method for quantitative transcriptional analysis of pathogens in vivo applied to the fungus Candida albicans[J].MBio,2015,6(5):e00942-915.
[6] Lucchini R,Sogo JM.Different chromatin structures along thespacers flanking active and inactive Xenopus rRNA genes[J].Mol Cell Biol,1992,12(10):4288-4296.
[7] Dammann R,Lucchini R,Koller T,et al.Transcription in the yeast rRNA gene locus:distribution of the active gene copies and chromatin structure of their flanking regulatory sequences[J].Mol Cell Biol,1995,15(10):5294-5303.
[8] Vanrobays E,Gleizes PE,Bousquet-Antonelli C,et al.Processing of 20S pre-rRNA to 18S ribosomal RNA in yeast requires Rrp10p,an essential non-ribosomal cytoplasmic protein[J].EMBO,2001,20(15):4204-4213.
[9] Keese CR,Bhawe K,Wegener J,et al.Real-time impedance assay to follow the invasive activities of metastatic cells in culture[J].Biotechniques,2002,33(4):842-844,846,848-850.
[10] Davydov DR.Microsomal monooxygenase in apoptosis:another target for cytochrome c signaling[J]? Trends Biochem Sci,2001,26(3),155-160.
[11] Zlotorynski E.Apoptosis:DR5 unfolds ER stress[J].Nat Rev Mol Cell Biol,2014,15(8):498-499.
[12] Sundar Rajan S,Srinivasan V,Balasubramanyam M.Endoplasmic reticulum(ER) stress & diabetes.[J].India Med Res,2007,125(3):411-424.
[13] Oka T,Ouchida M,Koyama M,et al.Gene silencing of the tyrosine phosphatase SHP1 gene by aberrant methylation in leukemias/lymphomas[J].Cancer Res,2002,62 (22):6390-6394.
[14] Andrew J.Phillips,Jonathan D,et al.Ras pathway signaling accelerates programmed cell death in the pathogenic fungus Candida albicans[J].PNAS January,2006,103(3):726-731.
[15] Keppetipola N,Shuman S.Characterization of the 2',3' cyclic phosphodiesterase activities of Clostridium thermocellum polynucleotide kinase-phosphatase and bacteriophage lambda phosphatase[J].Nucleic Acids Res,2007,35(22):7721-7732.
[16] Agrawal G,Joshi S,Subramani S.Cell-free sorting of peroxisomal membrane proteins from the endoplasmic reticulum[J].Proc Natl Acad Sci U S A,2011 108(22):9113-9118.
[17] Maaser K,H pfner M,Jansen A.Specific ligands of the peripheral benzodiazepine receptor induce apoptosis and cell cycle arrest in human colorectal cancer cells[J].Cancer,2001,85(11):1771-1780.
[18] Norlela S,Nor Azmi K,Khalid BA.Pituitary thyroid resistance syndrome[J].Med Malaysia,2005,60(5):642-643.
[19] Rusnak F,Mertz P.Calcineurin:form and function[J].Physiol Rev,2000,80(4):1483-1521.
[20] 徐臻.SET-TAF-I和CHMP6省略表达纯化以及初步晶体学分析[M].安徽:中国科学技术大学,生物化学与分子生物学,2010.
[21] Wollert T,Yang D,Ren X,et al.The ESCRT machinery at a glance[J].Cell Sci,2009,122(Ptl 3):2163-2166.