Abstract
Purpose: The effect of FK506 on endoplasmic reticulum(ER) derived calcium and caspase-12 mediated apoptosis in Jurkat human T-lymphocyte was investigated. Method: Cell viability was measured by flow cytometry. Intracellular calcium generation was measured. Western blotting of procaspase-12 was performed. And the catalytic activity of caspase -3, -6, -8, and -9 proteases in Jurkat cells was also measured. Results: FK506 dose-dependently decreased the viability in Jurkat cells. Increased intracellular accumulation of calcium in FK506 treated Jurkat cells from 24hours. FK506 continuously increased calcium concentration from 24hours to 72hrs. There was no evidence of calcium ionorpore (A23187) increased intracellular calcium changes with or without FK506 but calcium ATPase inhibitor (Thapsigargin) increased intracellular calcium accumulation and FK506 more and more increased calcium ATPase inhibitor(Thapsigargin) derived intracellular calcium accumulation. Procaspase-12 protease analyzed from 48hours. Treatment of cells with FK506 increased activation of caspase-12 protease. FK506 increased the catalytic activity of caspase-3 but there was no evidence of increased catalytic activation of caspase-6, -8 and -9 proteases in Jurkat cells. Conclusion: These data indicate that understanding of ER derived calcium and caspase-12 mediated apoptosis in human Jurkat cell line.
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