[Effects of histone deacetylase inhibitor MS-275 on expression of survivin and NF-kappaB in the human myeloma cell line U266 cells].
OBJECTIVE
To study the effects of HDACi on the expression of survivin and NF-kappaB in human myeloma cell line U266 cells.


METHODS
U266 cells were 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 changes were observed with Wright-Giemsa staining. The cell cycle was analyzed by flow cytometry, and the proteins of poly (ADP-ribose) polymerase (PARP), caspase-3, survivin, p21, CDK4 and IkappaB-alpha were detected by Western blotting.


RESULTS
MS-275 inhibited the growth of U266 cells in a dose- and time-dependent manner. The cell cycle was arrested at G(0)/G(1) phase. After exposure at 1.39 micromol/L MS-275 for 48 hours, the cell viability was decreased to 50%. The cell ratios of G(0)/G(1) phase were increased to (64.57 +/- 4.09)% for 24 h and (87.20 +/- 2.83)% for 36 h after 2 micromol/L MS-275 treatment. The visible morphological changes of U266 cells were confirmed with Wright-Giemsa staining. Cleaved-PARP, increased expression of p21, downregulation of expression of survivin, CDK4 and the phosphorylation of IkappaB-alpha was found by Western blot in MS-275 treated U266 cells.


CONCLUSIONS
MS-275-induced apoptosis of U266 cells is mediated by downregulation of expression of survivin and inactivation of NF-kappaB survival signalling pathways.
Ma
Jian
J
Department of Hematology of First Affiliated Hospital of General Hospital of PLA, Beijing 100037, China. mashixuan@163.com
Zhao
Ming
M
Yu
Xiao-Dan
XD
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; D002470:Cell Survival / Q000187:drug effects; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D056572:Histone Deacetylase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009101:Multiple Myeloma / Q000378:metabolism* / Q000473:pathology; D016328:NF-kappa B / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011725:Pyridines / Q000008:administration & dosage / Q000494:pharmacology*; D015398:Signal Transduction; D000077022:Survivin
C107489:BIRC5 protein, human; D001549:Benzamides; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D056572:Histone Deacetylase Inhibitors; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D016328:NF-kappa B; D011725:Pyridines; D000077022:Survivin; C118739:entinostat; D011065:Poly(ADP-ribose) Polymerases; D051358:Cyclin-Dependent Kinase 4; D053148:Caspase 3