[MS-275, a histone deacetylase inhibitor, induces apoptosis and alters survivin gene expression in human myeloma cell line U266].
OBJECTIVE
Histone deacetylase inhibitors (HDACi) exert antitumor effects through the induction of apoptosis. This study was to investigate the effects of HDACi on survivin gene expression, cell proliferation and apoptosis in human myeloma cell line U266.


METHODS
U266 cells were exposed to different concentrations of MS-275, a histone deacetylase inhibitor, at different time courses. Cell viability was measured using the trypan blue exclusion assay. Changes in cell morphology were observed with Wright-Giemsa staining. Cell cycle was analyzed using flow cytometry. Protein expressions of poly (ADP-ribose) polymerase (PARP), caspase-3, survivin, P21 and CDK4 were detected by Western blot.


RESULTS
MS-275 inhibited the growth of U266 cells in a dose-and time-dependent manner. After exposure to 1.39 micromol/L MS-275 for 48 h, the cell cycle was arrested at the G0/G1 phase, and the cell viability was decreased by 50%. After the treatment with 2 micromol/L MS-275 for 24 h and 36 h, the ratios of U266 cells at the G0/G1 phase were increased to 66.39% and 89.80%, respectively. Obvious changes in morphology of U266 were observed under microscopy. Cleaved PARP appeared, along with an increased protein level of P21 and a decrease of survivin and CDK4 levels in U266 cells treated with MS-275.


CONCLUSIONS
MS-275 could suppress the proliferation, induce apoptosis and reduce the expression of survivin in human myeloma cell line U266, which may be associated with the down-regulation of survivin.
Ma
Jian
J
Department of Hematology, First Affiliated Hospital of General Hospital of PLA, Beijing, China. mashixuan@163.com
Zhao
Ming
M
Yu
Xiao-Dan
XD
Wang
Zhi-Hong
ZH
D000107:Acetylation; D017209:Apoptosis / Q000187:drug effects*; D001549:Benzamides / Q000008:administration & dosage / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D015972:Gene Expression Regulation, Neoplastic; D056572:Histone Deacetylase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism*; D009101:Multiple Myeloma / Q000378:metabolism / Q000473:pathology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011725:Pyridines / Q000008:administration & dosage / Q000494:pharmacology*; D000077022:Survivin
C107489:BIRC5 protein, human; D001549:Benzamides; D050759:Cyclin-Dependent Kinase Inhibitor p21; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D051034:Inhibitor of Apoptosis Proteins; D011725:Pyridines; D000077022:Survivin; C118739:entinostat; D011065:Poly(ADP-ribose) Polymerases; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4; D053148:Caspase 3