Apoptosis Gene Expression Pattern Analysis of Jurkat Cells Treated with FK506

Tae Young Jang; Jae Sook Lee; Go Woon Woo; Hyun Chul Kim; Ho Kyun Lee; Sang Young Chung

doi:10.4174/jkss.2009.77.4.225

Journal List > J Korean Surg Soc > v.77(4) > 1011012

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Jang, Lee, Woo, Kim, Lee, and Chung: Apoptosis Gene Expression Pattern Analysis of Jurkat Cells Treated with FK506

Original Article

Journal of the Korean Surgical Society

Journal of the Korean Surgical Society 2009; 77(4): 225-237.

Published online: 30 September 2009

DOI: https://doi.org/10.4174/jkss.2009.77.4.225

Apoptosis Gene Expression Pattern Analysis of Jurkat Cells Treated with FK506

Tae Young Jang, M.D., Jae Sook Lee, M.D.¹, Go Woon Woo, M.D., Hyun Chul Kim, M.D., Ho Kyun Lee, M.D.², Sang Young Chung, M.D.²

Department of Surgery, Hanil Hospital, Seoul, Korea.

¹Korea Food & Drug Administration, Seoul, Korea.

²Department of Surgery, Chonnam National University Medical School, Gwangju, Korea.

Corresponding author (sycpvts@jnu.ac.kr)

Received 31 March 2009 Accepted 21 July 2009

Abstract

Purpose

FK506 (tacrolimus) is a widely used immunosuppressive agent in the treatment of various medical conditions, including autoimmune disease, bone marrow and organ transplantations. Previously FK506 was known to cause apoptotic death of human Jurkat T cells.

Methods

The current study was designed to analyze the gene expression pattern of Jurkat T cells after FK506 application by using cDNA microarray. Treatment of Jurkat T cells with FK506 resulted in a decrease of cell viability in a time- and dose-dependent manner. Next, total RNA of Jurkat T cells was extracted by using TRIzol reagent and used to carry out a confirmation test for the purity and integrity of total RNA.

Results

Gene expression levels related to apoptosis and cell cycle process were mainly focused to analyze in FK506-treated Jurkat T cells. According to the inhibition of calcineurin activity, MARCKS in PKC substrates and Sp3 transcription factor was markedly increased in FK506-treated cells. Also, cell cycle control gene Id1 and Id3 were induced in expression from FK506-treated Jurkat T cells. However, FK506 decreased the expression of Src homology 2, G protein, and MEK 2 genes in bioactive peptide induced signaling pathway. It also reduced the expression level of the insulin receptor, DRPLA and Bai1-associated protein 2 genes, which are involved in the regulation of cell motility and morphology control.

Conclusion

The author will continue to pursue the exact functional roles of genes that are markedly changed in expression by FK506 in human Jurkat T cells in vitro and in vivo experimental models.

Keywords: Tacrolimus, Gene expression, Apoptosis, Jurkat cell

Figures and Tables

Fig. 1

FK506 decreased the viability of Jurkat cells in dose- and time-dependent manners. Cells were treated with various concentration of FK506 for 96 hours, then cell viability was measured by MTT assay after FK506 treatment. Data represent the mean±standard deviation (SD) of triplicates.

Fig. 2

Modulation of gene expression following FK506-treated Jurkat cells. Total RNA was prepared from Jurkat using TRIzol as described by manufacturer. (A) Scatter plot analysis. The x and y axes represent Cy3 (Control) and Cy5 (FK506 10µM or 20µM) signal intensity values, respectively. These represent up-regulated (upper lines; red spots) or down-regulated (under lines; green spots) genes. (B) MA plot analysis. The x and y axes represent A (R+G/2) and M (R/G) signal intensity values. The R and G repesent Cy5-background and Cy3-background signal intensity, respectively. These represent up-regulated (upper lines; red spots) or down-regulated (under lines; green spots) genes.

Fig. 3

Cluster image showing the different classes of total gene expression profiles.

Table 1

RNA furification for microarray

^*RIN = RNA integrity number.

Table 2

Two-fold increase of biological processes

Table 3

Two-fold positive regulation of programmed cell death (7 Genes)

Table 4

Two-fold G1/S transition of mitotic cell cycle (3 Genes)

Table 5

Negative two-fold regulated biological process

Table 6

Negative two-fold regulated cell cycle process (32 Genes)

Table 7

Negative two-fold regulated cell death (33 Genes)

Table 8

Changes associated with calcineurin in keratinocyte differentiation by FK506

Table 9

Changes of Id proteins in regulating the G0-to-S transition of the cell cycle by FK506

Table 10

Changes of bioactive peptide induced signaling pathway by FK506

Table 11

Changes of gene associated with Diabetes and DRPLA pathway

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