title: Combinatorial treatment of DNA and chromatin-modifying drugs cause cell death in human and canine osteosarcoma cell lines.
abstract: Downregulation of microRNAs (miRNAs) at the 14q32 locus stabilizes the expression of cMYC, thus significantly contributing to osteosarcoma (OS) pathobiology. Here, we show that downregulation of 14q32 miRNAs is epigenetically regulated. The predicted promoter regions of miRNA clusters at 14q32 locus showed no recurrent patterns of differential methylation, but Saos2 cells showed elevated histone deacetylase (HDAC) activity. Treatment with 4-phenylbutyrate increased acetylation of histones associated with 14q32 miRNAs, but interestingly, robust restoration of 14q32 miRNA expression, attenuation of cMYC expression, and induction of apoptosis required concomitant treatment with 5-Azacytidine, an inhibitor of DNA methylation. These events were associated with genome-wide gene expression changes including induction of pro-apoptotic genes and downregulation of cell cycle genes. Comparable effects were achieved in human and canine OS cells using the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA/Vorinostat) and the DNA methylation inhibitor Zebularine (Zeb), with significantly more pronounced cytotoxicity in cells whose molecular phenotypes were indicative of aggressive biological behavior. These results suggested that the combination of these chromatin-modifying drugs may be a useful adjuvant in the treatment of rapidly progressive OS.
authors.affiliation: Department of Surgery, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Venugopal
authors.initials: V
authors.lastname: Thayanithy
authors.forename: ChangWon
authors.initials: C
authors.lastname: Park
authors.forename: Aaron L
authors.initials: AL
authors.lastname: Sarver
authors.forename: Reena V
authors.initials: RV
authors.lastname: Kartha
authors.forename: Derek M
authors.initials: DM
authors.lastname: Korpela
authors.forename: Ashley J
authors.initials: AJ
authors.lastname: Graef
authors.forename: Clifford J
authors.initials: CJ
authors.lastname: Steer
authors.forename: Jaime F
authors.initials: JF
authors.lastname: Modiano
authors.forename: Subbaya
authors.initials: S
authors.lastname: Subramanian
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D017209:Apoptosis; D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism*; D003562:Cytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D004247:DNA / Q000378:metabolism*; D019175:DNA Methylation; D004285:Dogs; D004354:Drug Screening Assays, Antitumor; D044127:Epigenesis, Genetic; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D012516:Osteosarcoma / Q000188:drug therapy*; D000077337:Vorinostat
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D002843:Chromatin; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D000077337:Vorinostat; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside; D004247:DNA