title: A unique histone deacetylase inhibitor alters microRNA expression and signal transduction in chemoresistant ovarian cancer cells.
abstract: Previously, we demonstrated potent antineoplastic activity of a distinctive histone deacetylase inhibitor (HDACI), AR42, against chemoresistant CP70 ovarian cancer cells in vitro and in vivo. Here, in follow-up to that work, we explored AR42 global mechanisms-of-action by examining drug-associated, genome-wide microRNA and mRNA expression profiles, which differed from those of the well-studied HDACI vorinostat. Expression of microRNA genes in negative correlation with their "target" coding gene (mRNA) transcripts, and transcription factor genes with expression positively correlated with coding genes having their cognate binding sites, were identified and subjected to gene ontology analyses. Those evaluations showed AR42 gene expression patterns to negatively correlate with Wnt signaling (> 18-fold induction of SFRP1), the epithelial-to-mesenchymal transition (40% decreased ATF1), and cell cycle progression (33-fold increased  14-3-3σ). By contrast, AR42 transcriptome alterations correlated positively with extrinsic ("death receptor") apoptosis (> 2.3-fold upregulated DAPK) and favorable ovarian cancer histopathology and prognosis. Inhibition of Wnt signaling was experimentally validated by: (1) > 2.6-fold reduced Wnt reporter activity; and (2) 36% reduction in nuclear, activated β-catenin. Likely AR42 induction of multiple (type I or type II autophagic) cell death cascades was further supported by 57% decreased reliance upon reactive oxygen, increased mitochondrial membrane disruption, and caspase independence, as compared with vorinostat. Taken together, we demonstrate distinct antineoplastic pathway alterations, in aggressive ovarian cancer cells, following treatment with a promising HDACI, AR42. These combined computational and experimental approaches may also represent a straightforward means for mechanistic studies of other promising antineoplastics, and/or the identification of agents that may complement epigenetic therapies.
authors.affiliation: Medical Sciences Program, Indiana University, Bloomington, IN, USA. rbalch@indiana.edu
authors.forename: Curt
authors.initials: C
authors.lastname: Balch
authors.forename: Kaleb
authors.initials: K
authors.lastname: Naegeli
authors.forename: Seungyoon
authors.initials: S
authors.lastname: Nam
authors.forename: Brett
authors.initials: B
authors.lastname: Ballard
authors.forename: Alan
authors.initials: A
authors.lastname: Hyslop
authors.forename: Christina
authors.initials: C
authors.lastname: Melki
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Reilly
authors.forename: Man-Wook
authors.initials: MW
authors.lastname: Hur
authors.forename: Kenneth P
authors.initials: KP
authors.lastname: Nephew
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D019008:Drug Resistance, Neoplasm; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D058977:Molecular Sequence Annotation; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism*; D010654:Phenylbutyrates / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D051153:Wnt Proteins / Q000378:metabolism
chemical_list: D056572:Histone Deacetylase Inhibitors; D035683:MicroRNAs; C529674:OSU-HDAC42 compound; D010654:Phenylbutyrates; D012333:RNA, Messenger; D051153:Wnt Proteins