Transcriptional Profiles Of Trichophyton Rubrum Under The Treatment Of5-flucytosine, Cyclosporine A And FK506

Fungus is an important pathogen besides of bacterium and virus to evoke human chronic infectious disease, which can cause superficial and deep mycosis, with high contagiousness and hard to cure. Trichophyton rubrum is one of the most popular etiological agents among dermatophyte, it can provoke superficial part disease, for example, tinea capitis, onychomycosis, ringworm of feet, and it can also produce deep mycotic infection, even endanger to life. To be eukaryote, fungi has some similar structure with the cell of human being, therefore, toxic effects, adverse reaction, resistance and cross tolerance were usually observed during the treatment of antifungal agents in clinical treatment. How to find safe, effective drug target and how to develop broad-spectrum, low poisonous, high performance and no cross tolerance antifungal drugs become a significant task for mycology.In the techniques of screening potential drug targets, gene chip technology is predominant, which can work by large scale, high-flux, parallel, and can analyze the mechanism of action, medicine activity, and drug toxicity on gene level. We had constructed the Expressed Sequence Tag (EST) database of Trichophyton rubrum before, from which we manufactured cDNA chip of8,997EST (altogether1,003control, including blank control, negative control and positive control), each chip hold10,000dots. We applied the cDNA chip to research the transcriptional profiles for Trichophyton rubrum under the treatment of5-flucytosine, cyclosporine A and FK506respectively.We got the first transcriptional profiles of5-flucytosine on Trichophyton rubrum. 474genes were differentially expressed, among them,196genes were up-regulated, and278genes were down-regulated. Gene CDC21involved in nucleotide transport and metabolism, were differentially expressed, which confirmed that the mechanism of action of5-flucytosine is depressing the activity of thymidylate synthase and restraining the biosynthesis of DNA, RNA and nuclear division. The newly found gene involved in transcription, E2F1, might be the potential antifungal target. Gene YCF1and SNQ2might connect with the potential mechanism of action about the drug resistance for5-flucytosine.We got the first transcriptional profiles of cyclosporine A on Trichophyton rubrum.410genes were differentially expressed, among them,228genes were up-regulated, and182genes were down-regulated. Genes involved in signal transduction mechanisms, such as DW686139,DW705207, involved in inorganic ion transport and metabolism, such as DW692468,DW697092, involved in Cell cycle control, cell division, chromosome partitioning, such as BIK1,CDC25C and KIN4, were differentially expressed. They validated the mechanism of action for cyclosporine A is combining with immunophilin Cyclophilin to inhibit the activity of calcineurin and peptidyl-prolyl cis-trans isomerase. Gene involved in lipid transport and metabolism, ERG2, was down-regulated, this proved the report that better curative effect were achieved when cyclosporine A combined with antifungal agents which target the lipid synthesis. We suggested that gene involved in translation, ribosomal structure and biogenesis, DW705297encodes isoleucyl-tRNA synthetase, might be the possible antifungal target.We got the first transcriptional profiles of FK506on Trichophyton rubrum,549 genes were differentially expressed, among them,348genes were up-regulated, and201genes were down-regulated. Genes involved in signal transduction mechanisms, such as DW686139, DW705207, MAPK, MKH1, involved in Cell wall/membrane/envelope biogenesis, such as DW704622, DW699144, involved in posttranslational modification, protein turnover, chaperones, such as DW685744, and involved in amino acid transport and metabolism, such as DW698808, EL790430, were differentially expressed, which verified the mechanism of action of FK506is restraining the activity of calcineurin. Gene involved in amino acid transport and metabolism, MET6, might has synergism interaction with ergosterol and calcineurin inhibitor, we proposed that MET6might be the potential antifungal target.Real-time quantitative reverse transcriptase polymerase chain reaction technique was utilized to certify the differentially expressed genes selected from the treatment of5-flucytosine, cyclosporine A and FK506on Trichophyton rubrum respectively, the result showed that the outcome of chip experiment is reliable.Our research testified the known mechanism of action of5-flucytosine, cyclosporine A and FK506, found some new genes, and gave some cue for the potential drug target, potential new mechanism of action and potential mechanism of drug resistance, which will offer useful information for the development of new antifungal agents.