Anisomycin suppresses Jurkat T cell growth by the cell cycle-regulating proteins

Background: recent studies have shown that anisomycin significantly inhibits mammalian cell proliferation,but its mechanism remains unclear. in this study,jurkat t cells were used to first explore a relationship between effect of anisomycin on them and alteration of cell cycle-regulating proteins. methods: cell colony formation,cck-8 assay,flow cytometry,rt-pcr and western blot were employed to evaluate correlation of ten cell cycle-regulating proteins with suppression of the cell proliferation and arrest of the cell cycle by anisomycin. results: our data showed that anisomycin inhibited the colony-formation and proliferation of jurkat t cells in a dose-dependent manner,and arrested the cells into s and g2/m phases with the production of sub-diploid cells. the levels of p21,p-p27 and p53/p-p53 reached their peaks 4 h after anisomycin treatment,presenting a positive correlation with anisomycin concentration,and p16,p-p21,p27,p57,p73/p-p73 and p-rb changed little with the prolonged exposure time or increased concentrations of anisomycin. but the level of rb protein was increased at 24 h after the treatment of anisomycin. the expression of an inverted ccaat box binding protein (icbp90) in jurkat t cells came to decrease 12 h after the treatment of anisomycin,presenting a negative correlation with anisomycin concentration. subsequently,the expression of p-cdk2 was also decreased at 24 h,presenting an obviously negative correlation,whereas p-cdk1 showed no differences among the differently treated jurkat t cells. furthermore,the level of p21 and p53 mrna was increased with the enhanced concentrations of anisomycin. conclusion: the results indicate that anisomycin may activate the p53/p21/p27 signaling to decrease the expression of icbp90,inhibit expression of p-cdk2 to block the cells into s and g2/m phases,and finally result in proliferation inhibition of jurkat t cells. copyright © 2013 by institute of pharmacology.