title: The intestinal epithelial cell differentiation marker intestinal alkaline phosphatase (ALPi) is selectively induced by histone deacetylase inhibitors (HDACi) in colon cancer cells in a Kruppel-like factor 5 (KLF5)-dependent manner.
abstract: The histone deacetylase inhibitor (HDACi) sodium butyrate promotes differentiation of colon cancer cells as evidenced by induced expression and enzyme activity of the differentiation marker intestinal alkaline phosphatase (ALPi). Screening of a panel of 33 colon cancer cell lines identified cell lines sensitive (42%) and resistant (58%) to butyrate induction of ALP activity. This differential sensitivity was similarly evident following treatment with the structurally distinct HDACi, MS-275. Resistant cell lines were significantly enriched for those harboring the CpG island methylator phenotype (p = 0.036, Chi square test), and resistant cell lines harbored methylation of the ALPi promoter, particularly of a CpG site within a critical KLF/Sp regulatory element required for butyrate induction of ALPi promoter activity. However, butyrate induction of an exogenous ALPi promoter-reporter paralleled up-regulation of endogenous ALPi expression across the cell lines, suggesting the presence or absence of a key transcriptional regulator is the major determinant of ALPi induction. Through microarray profiling of sensitive and resistant cell lines, we identified KLF5 to be both basally more highly expressed as well as preferentially induced by butyrate in sensitive cell lines. KLF5 overexpression induced ALPi promoter-reporter activity in resistant cell lines, KLF5 knockdown attenuated butyrate induction of ALPi expression in sensitive lines, and butyrate selectively enhanced KLF5 binding to the ALPi promoter in sensitive cells. These findings demonstrate that butyrate induction of the cell differentiation marker ALPi is mediated through KLF5 and identifies subsets of colon cancer cell lines responsive and refractory to this effect.
authors.affiliation: From the Department of Oncology, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, New York 10461.
authors.forename: Joongho
authors.initials: J
authors.lastname: Shin
authors.affiliation: From the Department of Oncology, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, New York 10461.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Carr
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Georgia A
authors.initials: GA
authors.lastname: Corner
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Lars
authors.initials: L
authors.lastname: Tögel
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Mercedes
authors.initials: M
authors.lastname: Dávalos-Salas
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Mercedes
authors.initials: M
authors.lastname: Dávaos-Salas
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Hoanh
authors.initials: H
authors.lastname: Tran
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Anderly C
authors.initials: AC
authors.lastname: Chueh
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Sheren
authors.initials: S
authors.lastname: Al-Obaidi
authors.affiliation: the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia.
authors.forename: Fiona
authors.initials: F
authors.lastname: Chionh
authors.affiliation: From the Department of Oncology, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, New York 10461.
authors.forename: Naseem
authors.initials: N
authors.lastname: Ahmed
authors.affiliation: the Queensland Institute of Medical Research, 300 Herston Road, Herston, Queensland 4006, Australia.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Buchanan
authors.affiliation: the Queensland Institute of Medical Research, 300 Herston Road, Herston, Queensland 4006, Australia.
authors.forename: Joanne P
authors.initials: JP
authors.lastname: Young
authors.affiliation: the Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Madhu S
authors.initials: MS
authors.lastname: Malo
authors.affiliation: the Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Hodin
authors.affiliation: the Group of Molecular Oncology, Centro en Investigación en Bioquímica y Biología Molecular-Nanomedicine, Vall d'Hebron University Hospital, Research Institute, Universitat Autònoma de Barcelona, Passeig Vall d'Hebron, 119-129, 08035 Barcelona, Spain and El Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina, Spain.
authors.forename: Diego
authors.initials: D
authors.lastname: Arango
authors.affiliation: the Walter and Eliza Hall Institute, Melbourne 3052, Australia.
authors.forename: Oliver M
authors.initials: OM
authors.lastname: Sieber
authors.affiliation: From the Department of Oncology, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, New York 10461.
authors.forename: Leonard H
authors.initials: LH
authors.lastname: Augenlicht
authors.affiliation: Peter MacCallum Cancer Center, Melbourne 3002, Australia, and.
authors.forename: Amardeep S
authors.initials: AS
authors.lastname: Dhillon
authors.affiliation: the Veterans Affairs New York Harbor Health Care System, Brooklyn, New York 11209.
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Weber
authors.affiliation: From the Department of Oncology, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, New York 10461, the Ludwig Institute for Cancer Research, Austin Health, Melbourne 3084, Australia, john.mariadason@ludwig.edu.au.
authors.forename: John M
authors.initials: JM
authors.lastname: Mariadason
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism*; D001549:Benzamides / Q000494:pharmacology; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D020148:Butyric Acid / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism*; D020869:Gene Expression Profiling; D045325:HCT116 Cells; D019073:HT29 Cells; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011725:Pyridines / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D001549:Benzamides; D056572:Histone Deacetylase Inhibitors; C492214:KLF5 protein, human; D051741:Kruppel-Like Transcription Factors; D011725:Pyridines; D020148:Butyric Acid; C118739:entinostat; D000469:Alkaline Phosphatase
keywords: CIMP; Colon Cancer; DNA Methylation; Differentiation; HDAC; Histone Deacetylase Inhibitor (HDAC Inhibitor) (HDI); Kruppel-like Factor (KLF)