title: Reciprocal binding of CTCF and BORIS to the NY-ESO-1 promoter coincides with derepression of this cancer-testis gene in lung cancer cells.
abstract: Regulatory sequences recognized by the unique pair of paralogous factors, CTCF and BORIS, have been implicated in epigenetic regulation of imprinting and X chromosome inactivation. Lung cancers exhibit genome-wide demethylation associated with derepression of a specific class of genes encoding cancer-testis (CT) antigens such as NY-ESO-1. CT genes are normally expressed in BORIS-positive male germ cells deficient in CTCF and meCpG contents, but are strictly silenced in somatic cells. The present study was undertaken to ascertain if aberrant activation of BORIS contributes to derepression of NY-ESO-1 during pulmonary carcinogenesis. Preliminary experiments indicated that NY-ESO-1 expression coincided with derepression of BORIS in cultured lung cancer cells. Quantitative reverse transcription-PCR analysis revealed robust, coincident induction of BORIS and NY-ESO-1 expression in lung cancer cells, but not normal human bronchial epithelial cells following 5-aza-2'-deoxycytidine (5-azadC), Depsipeptide FK228 (DP), or sequential 5-azadC/DP exposure under clinically relevant conditions. Bisulfite sequencing, methylation-specific PCR, and chromatin immunoprecipitation (ChIP) experiments showed that induction of BORIS coincided with direct modulation of chromatin structure within a CpG island in the 5'-flanking noncoding region of this gene. Cotransfection experiments using promoter-reporter constructs confirmed that BORIS modulates NY-ESO-1 expression in lung cancer cells. Gel shift and ChIP experiments revealed a novel CTCF/BORIS-binding site in the NY-ESO-1 promoter, which unlike such sites in the H19-imprinting control region and X chromosome, is insensitive to CpG methylation in vitro. In vivo occupancy of this site by CTCF was associated with silencing of the NY-ESO-1 promoter, whereas switching from CTCF to BORIS occupancy coincided with derepression of NY-ESO-1. Collectively, these data indicate that reciprocal binding of CTCF and BORIS to the NY-ESO-1 promoter mediates epigenetic regulation of this CT gene in lung cancer cells, and suggest that induction of BORIS may be a novel strategy to augment immunogenicity of pulmonary carcinomas.
authors.affiliation: Thoracic Oncology Section, Surgery Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892-1201, USA.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Hong
authors.forename: Yang
authors.initials: Y
authors.lastname: Kang
authors.forename: Ziedulla
authors.initials: Z
authors.lastname: Abdullaev
authors.forename: Patrick T
authors.initials: PT
authors.lastname: Flanagan
authors.forename: Svetlana D
authors.initials: SD
authors.lastname: Pack
authors.forename: Maria R
authors.initials: MR
authors.lastname: Fischette
authors.forename: Mina T
authors.initials: MT
authors.lastname: Adnani
authors.forename: Dmitri I
authors.initials: DI
authors.lastname: Loukinov
authors.forename: Sergei
authors.initials: S
authors.lastname: Vatolin
authors.forename: John I
authors.initials: JI
authors.lastname: Risinger
authors.forename: Mary
authors.initials: M
authors.lastname: Custer
authors.forename: G Aaron
authors.initials: GA
authors.lastname: Chen
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.forename: Dao M
authors.initials: DM
authors.lastname: Nguyen
authors.forename: J Carl
authors.initials: JC
authors.lastname: Barrett
authors.forename: Victor V
authors.initials: VV
authors.lastname: Lobanenkov
authors.forename: David S
authors.initials: DS
authors.lastname: Schrump
mesh_terms: D000951:Antigens, Neoplasm / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D000076246:CCCTC-Binding Factor; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006657:Histones / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013447:Sulfites / Q000494:pharmacology
chemical_list: D000951:Antigens, Neoplasm; D000076246:CCCTC-Binding Factor; C104948:CTAG1B protein, human; C000619697:CTCF protein, human; C456540:CTCFL protein, human; D004268:DNA-Binding Proteins; D006657:Histones; D008565:Membrane Proteins; D012097:Repressor Proteins; D013447:Sulfites; C042345:hydrogen sulfite