Effects of histone deacetylase inhibitor MS-275 in vitro on apoptosis of U266 cell line
AIM: To study the effect of MS-275,an inhibitor of histone deacetylase,on the growth and apoptosis of myeloma cell line U266. METHODS: U266 cell was cultured in RPMI-1640 in the presence of MS-275 and the cell viability was evaluated by Trypan blue exclusion assay and cell count. The cell morphological change was observed with Wright-Giemsa staining. The cell cycle was analyzed by flow cytometry and the proteins of poly ( ADP-ribose ) polymerase ( PARP ),Caspase-3,Caspase-8 and Caspase-9 were detected by Western Blot. RESULTS: MS-275 inhibited the growth of U266 cells in a dose-and time-dependent manner. The cell cycle was arrested at G0/G1 phase. After exposure to 1. 39 μmol/L MS-275 for 48 h,the cell viability decreased to 50%. After 2 μmol/L MS-275 treatment for 24 h and 36 h,the cell ratios of G0/G1 phase increased respectively to 64. 57% and 87.20%. The morphological visible changes of U266 were confirmed by Wright-Giemsa staining. The proteins were abstracted from the treated cells,and Caspase-3,Caspase-8 and Caspase-9 activation and cleavage of PARP were examined by Western blot using a specific antibody. The cleaved Caspase-3,Caspase-8 and Caspase-9 and cleaved PARP were recognized respectively. CONCLUSION: MS-275 inhibits the proliferation and arrests the cell cycle at G0/G1 phase. MS-275 induces apoptosis of U266 cells through both extracellular and intracellular signaling cascade pathways.
histone deacetylase inhibitorsmultiple myelomacell line U266MS-275apoptosis
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