J Korean Surg Soc.  2013 Oct;85(4):161-168. 10.4174/jkss.2013.85.4.161.

MicroRNA profiling of tacrolimus-stimulated Jurkat human T lympocytes

• Lee HK
• Chung SY
• Na Choi SJ

• KMID: 2096706
• DOI: http://doi.org/10.4174/jkss.2013.85.4.161

Abstract

PURPOSE
This study investigated the Jurkat T cell line expresses cytotoxicity when treated with different concentrations of FK506, and analyzed the expression pattern of microRNA when stimulated by FK506 using the microRNAs microarray, as well as the expression pattern of a gene that is related to the differentiation, activation and proliferation of T cells after being affected by the change of microRNAs.
METHODS
To investigate the effects of FK506 on microRNA expression, we purified total RNA of Jurkat cells treated with 20 microM FK506 for 72 hours and used to analyze microRNA profiling by using Agilent's chip.
RESULTS
These results demonstrated that treatment with FK506 markedly induced the down-regulation of 20 microRNAs as well as the up-regulation of 20 microRNAs in a time-dependent manner. The genes that down-regulated by FK506 include let-7a*, miR-20a*, and miR-487a. Otherwise miR-202, miR-485-5p, and miR-518c* are gradually up-regulated in expression. Sanger Institute and DAVIDs bioinformatics indicated that microRNAs regulated the several transcriptomes including nuclear factor of activated T cell-related, T cell receptor/interleukin-2 signaling, and Ca(2+)-calmodulin-dependent phosphatase calcineurin pathways.
CONCLUSION
As a result of treating FK506 to a Jurkat cell line and running the microRNA microarray, it was found that FK506 not only took part in the suppression of T cell proliferation/activation by inhibiting calcineurin in Jurkat apoptosis, but also affected the microRNAs that are involved in the regulation of various signal transduction pathways.

Keyword

Tacrolimus; MicroRNAs; Jurkat T lymphocyte

MeSH Terms

Apoptosis
Calcineurin
Cell Line
Computational Biology
Down-Regulation
Genes, vif
Humans
Jurkat Cells
MicroRNAs
RNA
Running
Signal Transduction
T-Lymphocytes
Tacrolimus
Transcriptome
Up-Regulation
Calcineurin
MicroRNAs
RNA
Tacrolimus

Figure

• 

  Fig. 1 Regulation of microRNAs expression in Jurkat T cell after FK506 treatment. Differentially expressed microRNAs were analyzed by hierarchical clustering of the log 2 value of each microRNA microarray signal at 24, 48, and 72 hours. Red, up-regulation; Green, down-regulation; Black, no change.

• 

  Fig. 2 The microRNAs down regulated in the time dependent manners by FK506 treatment.

• 

  Fig. 3 The microRNAs up regulated in time dependent manner during FK506 treatments.

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