title: ICI182,780 induces p21Waf1 gene transcription through releasing histone deacetylase 1 and estrogen receptor alpha from Sp1 sites to induce cell cycle arrest in MCF-7 breast cancer cell line.
abstract: We used the estrogen-responsive MCF-7 breast cancer cell line as a relevant model to study the anti-proliferative effects of ICI182,780 and identified the negative cell cycle regulator p21Waf1 as a specific target of ICI182,780. Furthermore, silencing of the p21Waf1 expression by small interfering RNA overcame the G0/G1 cell cycle arrest induced by ICI182,780, suggesting that the induction of p21Waf1 expression has a direct role in mediating the ICI182,780-induced G0/G1 arrest. We further demonstrated that the induction of p21Waf1 by ICI182,780 is mediated at transcriptional and gene promoter levels through the proximal Sp1 sites located near the transcription start site. Co-immunoprecipitation, DNA "pull-down," and chromatin immunoprecipitation experiments together showed that in cycling cells, estrogen receptor alpha and histone deacetylase 1 (HDAC1) are recruited to the proximal Sp1 sites of the promoter to repress p21Waf1 expression. In the presence of ICI182,780, estrogen receptor alpha and HDACs are dissociated from Sp1, resulting in increased histone acetylation and de-repression of the p21Waf1 promoter and induction of p21Waf1 expression. The fact that p21Waf1 expression is normally repressed by HDAC activity in cycling cells is further demonstrated by the finding that p21Waf1 transcription can be induced by the silencing of HDACs with small interfering RNA or treatment with HDAC inhibitors.
authors.affiliation: Cancer Research-UK Laboratories and Section of Cancer Cell Biology, Department of Cancer Medicine, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 ONN, United Kingdom.
authors.forename: Rana
authors.initials: R
authors.lastname: Varshochi
authors.forename: Faezah
authors.initials: F
authors.lastname: Halim
authors.forename: Andrew
authors.initials: A
authors.lastname: Sunters
authors.forename: John P
authors.initials: JP
authors.lastname: Alao
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Madureira
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Hart
authors.forename: Simak
authors.initials: S
authors.lastname: Ali
authors.forename: David M
authors.initials: DM
authors.lastname: Vigushin
authors.forename: R Charles
authors.initials: RC
authors.lastname: Coombes
authors.forename: Eric W-F
authors.initials: EW
authors.lastname: Lam
mesh_terms: D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology*; D001665:Binding Sites; D001943:Breast Neoplasms; D018797:Cell Cycle Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D050759:Cyclin-Dependent Kinase Inhibitor p21; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology*; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D000077267:Fulvestrant; D016193:G1 Phase / Q000187:drug effects; D020868:Gene Silencing; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000502:physiology; D034741:RNA, Small Interfering; D016329:Sp1 Transcription Factor / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: D018931:Antineoplastic Agents, Hormonal; C493956:CDKN1A protein, human; D018797:Cell Cycle Proteins; D050759:Cyclin-Dependent Kinase Inhibitor p21; D004791:Enzyme Inhibitors; D047628:Estrogen Receptor alpha; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D034741:RNA, Small Interfering; D016329:Sp1 Transcription Factor; D000077267:Fulvestrant; C012589:trichostatin A; D004958:Estradiol; D006655:Histone Deacetylases