title: Transcriptional targets of the chromatin-remodelling factor SMARCA4/BRG1 in lung cancer cells.
abstract: BRG1, also called SMARCA4, is the catalytic subunit of the SWI/SNF chromatin-remodelling complex and influences transcriptional regulation by disrupting histone-DNA contacts in an ATP-dependent manner. BRG1 and other members of the SWI/SNF complex become inactivated in tumours, implying a role in cancer development. To understand the contribution of BRG1 to lung tumourigenesis, we restored BRG1 in H1299 lung cancer cells and used cDNA microarray analysis to identify changes in gene expression. Forty-three transcripts became activated, whereas two were repressed. Chromatin immunoprecipitation of resulting candidate genes revealed that the CYP3A4 and ZNF185 promoters recruited BRG1 and that recruitment to the CYP3A4 promoter was specific to this gene and did not involve the CYP3A5 or CYP3A7 family members. Moreover, specifically BRG1 but not its homologue BRM was recruited to the CYP3A4 and ZNF185 promoters. To explore their potential relevance in lung tumours, levels of CYP3A4 and ZNF185 transcripts were evaluated in seven additional lung cancer cell lines. CYP3A4 was undetectable in any of the lung cancer cells tested, and only the CYP3A5 family member was present in the A549 and Calu-3 cells. In contrast, the amount of ZNF185 transcript clearly varied among lung cancer cell lines and severely reduced levels were observed in BRG1-deficient cells, except those of A427. We extended these observations to 27 lung primary tumours using real-time RT-PCR (TaqMan) and observed that four (15%) and 14 (52%) of them had BRG1 and ZNF185 downregulation, respectively, when compared with normal lung. No significant correlation between reduced levels of BRG1 and ZNF185 was observed, indicating that additional mechanisms to BRG1 inactivation may contribute to the loss of ZNF185 expression in lung tumours. In conclusion, our results provide evidence that transcriptional activation of ZNF185 and CYP3A4 is mediated by direct association of BRG1 with their promoters and also indicate that a decreased level of ZNF185 is a common feature of lung tumours and may be of biological relevance in lung carcinogenesis.
authors.affiliation: Lung Cancer Group, Molecular Pathology Programme, Spanish National Cancer Centre, C/Melchior Fernandez Almagro 3, 28029 Madrid, Spain.
authors.forename: Pedro P
authors.initials: PP
authors.lastname: Medina
authors.forename: Julian
authors.initials: J
authors.lastname: Carretero
authors.forename: Esteban
authors.initials: E
authors.lastname: Ballestar
authors.forename: Barbara
authors.initials: B
authors.lastname: Angulo
authors.forename: Fernando
authors.initials: F
authors.lastname: Lopez-Rios
authors.forename: Manel
authors.initials: M
authors.lastname: Esteller
authors.forename: Montserrat
authors.initials: M
authors.lastname: Sanchez-Cespedes
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002843:Chromatin / Q000235:genetics*; D047369:Chromatin Immunoprecipitation; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D003598:Cytoskeletal Proteins; D004247:DNA / Q000378:metabolism; D004265:DNA Helicases; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D005819:Genetic Markers; D005822:Genetic Vectors; D006657:Histones / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D060588:LIM Domain Proteins; D008175:Lung Neoplasms / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D012150:Polymorphism, Restriction Fragment Length; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection
chemical_list: D002843:Chromatin; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D005819:Genetic Markers; D006657:Histones; D060588:LIM Domain Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; C108627:ZNF185 protein, human; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; C104464:CYP3A protein, human; C510164:CYP3A5 protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; C084108:SMARCA4 protein, human; D004265:DNA Helicases