Epigenetic inactivation of the potential tumor suppressor gene FOXF1 in breast cancer.

The expression of several members of the FOX gene family is known to be altered in a variety of cancers. We show that in breast cancer, FOXF1 gene is a target of epigenetic inactivation and that its gene product exhibits tumor-suppressive properties. Loss or downregulation of FOXF1 expression is associated with FOXF1 promoter hypermethylation in breast cancer cell lines and in invasive ductal carcinomas. Methylation of FOXF1 in invasive ductal carcinoma (37.6% of 117 cases) correlated with high tumor grade. Pharmacologic unmasking of epigenetic silencing in breast cancer cells restored FOXF1 expression. Re-expression of FOXF1 in breast cancer cells with epigenetically silenced FOXF1 genes led to G(1) arrest concurrent with or without apoptosis to suppress both in vitro cell growth and in vivo tumor formation. FOXF1-induced G(1) arrest resulted from a blockage at G(1)-S transition of the cell cycle through inhibition of the CDK2-RB-E2F cascade. Small interfering RNA-mediated depletion of FOXF1 in breast cancer cells led to increased DNA re-replication, suggesting that FOXF1 is required for maintaining the stringency of DNA replication and genomic stability. Furthermore, expression profiling of cell cycle regulatory genes showed that abrogation of FOXF1 function resulted in increased expression of E2F-induced genes involved in promoting the progression of S and G(2) phases. Therefore, our studies have identified FOXF1 as a potential tumor suppressor gene that is epigenetically silenced in breast cancer, which plays an essential role in regulating cell cycle progression to maintain genomic stability.

[1]  J. Darnell,et al.  Hepatocyte nuclear factor 3/fork head or "winged helix" proteins: a family of transcription factors of diverse biologic function. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[2]  S. Weitzman,et al.  Culture of normal and malignant primary human mammary epithelial cells in a physiological manner simulates in vivo growth patterns and allows discrimination of cell type. , 1993, Cancer research.

[3]  M. Groudine,et al.  Cloning of the human homolog of the CDC34 cell cycle gene by complementation in yeast. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[4]  J. Labbé,et al.  The MO15 gene encodes the catalytic subunit of a protein kinase that activates cdc2 and other cyclin‐dependent kinases (CDKs) through phosphorylation of Thr161 and its homologues. , 1993, The EMBO journal.

[5]  W. Sellers,et al.  A potent transrepression domain in the retinoblastoma protein induces a cell cycle arrest when bound to E2F sites. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[6]  P. Carlsson,et al.  Differential Activation of Lung-specific Genes by Two Forkhead Proteins, FREAC-1 and FREAC-2 (*) , 1996, The Journal of Biological Chemistry.

[7]  J. Herman,et al.  Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Johnston,et al.  Neuronal Cdc2-like Kinase (Nclk) Binds and Phosphorylates the Retinoblastoma Protein* , 1997, The Journal of Biological Chemistry.

[9]  David I. Smith,et al.  Role for the p53 homologue p73 in E2F-1-induced apoptosis , 2000, Nature.

[10]  Mingjie Zhang,et al.  Identification of a Common Protein Association Region in the Neuronal Cdk5 Activator* , 2000, The Journal of Biological Chemistry.

[11]  J. Welsh,et al.  Analysis of gene expression identifies candidate markers and pharmacological targets in prostate cancer. , 2001, Cancer research.

[12]  E. Lander,et al.  Classification of human lung carcinomas by mRNA expression profiling reveals distinct adenocarcinoma subclasses , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[13]  P. Carlsson,et al.  Haploinsufficiency of the forkhead gene Foxf1, a target for sonic hedgehog signaling, causes lung and foregut malformations. , 2001, Development.

[14]  R. Spang,et al.  Role for E2F in Control of Both DNA Replication and Mitotic Functions as Revealed from DNA Microarray Analysis , 2001, Molecular and Cellular Biology.

[15]  P. Carlsson,et al.  The forkhead transcription factor Foxf1 is required for differentiation of extra-embryonic and lateral plate mesoderm. , 2001, Development.

[16]  David E. Misek,et al.  Gene-expression profiles predict survival of patients with lung adenocarcinoma , 2002, Nature Medicine.

[17]  Yan Zhou,et al.  Haploinsufficiency of the Mouse Forkhead Box f1 Gene Causes Defects in Gall Bladder Development* , 2002, The Journal of Biological Chemistry.

[18]  P. Carlsson,et al.  Forkhead transcription factors: key players in development and metabolism. , 2002, Developmental biology.

[19]  J. Cheville,et al.  Transcriptional silencing of zinc finger protein 185 identified by expression profiling is associated with prostate cancer progression. , 2003, Cancer research.

[20]  Yan Zhou,et al.  Foxf1 +/− mice exhibit defective stellate cell activation and abnormal liver regeneration following CCl4 injury , 2003, Hepatology.

[21]  Avrum Spira,et al.  Gene expression in lung adenocarcinomas of smokers and nonsmokers. , 2003, American journal of respiratory cell and molecular biology.

[22]  S. Kearsey,et al.  Enigmatic variations: divergent modes of regulating eukaryotic DNA replication. , 2003, Molecular cell.

[23]  Jeffrey R Marks,et al.  Gene Expression Patterns That Characterize Advanced Stage Serous Ovarian Cancers , 2004, The Journal of the Society for Gynecologic Investigation: JSGI.

[24]  A. Datta,et al.  Foxm1b transcription factor is essential for development of hepatocellular carcinomas and is negatively regulated by the p19ARF tumor suppressor. , 2004, Genes & development.

[25]  A. Chinnaiyan,et al.  Integration of high-resolution array comparative genomic hybridization analysis of chromosome 16q with expression array data refines common regions of loss at 16q23–qter and identifies underlying candidate tumor suppressor genes in prostate cancer , 2004, Oncogene.

[26]  M. Katoh,et al.  Human FOX gene family (Review). , 2004, International journal of oncology.

[27]  David Botstein,et al.  Different gene expression patterns in invasive lobular and ductal carcinomas of the breast. , 2004, Molecular biology of the cell.

[28]  R. Tibshirani,et al.  Gene expression profiling identifies clinically relevant subtypes of prostate cancer. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[29]  E. Gelfand,et al.  Cyclin-dependent kinase 6 inhibits proliferation of human mammary epithelial cells. , 2004, Molecular cancer research : MCR.

[30]  H. Nishitani,et al.  DNA replication licensing. , 2004, Frontiers in bioscience : a journal and virtual library.

[31]  M. Becich,et al.  Gene expression alterations in prostate cancer predicting tumor aggression and preceding development of malignancy. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[32]  Mogens Kruhøffer,et al.  Gene Expression in the Urinary Bladder , 2004, Cancer Research.

[33]  Jiri Bartek,et al.  Checking on DNA damage in S phase , 2004, Nature Reviews Molecular Cell Biology.

[34]  M. P. Holloway,et al.  Aberrant Regulation of Survivin by the RB/E2F Family of Proteins* , 2004, Journal of Biological Chemistry.

[35]  Shinichiro Wachi,et al.  Interactome-transcriptome analysis reveals the high centrality of genes differentially expressed in lung cancer tissues , 2005, Bioinform..

[36]  Clifford A. Meyer,et al.  Chromosome-Wide Mapping of Estrogen Receptor Binding Reveals Long-Range Regulation Requiring the Forkhead Protein FoxA1 , 2005, Cell.

[37]  J. Julian Blow,et al.  Preventing re-replication of chromosomal DNA , 2005, Nature Reviews Molecular Cell Biology.

[38]  Raphael A Nemenoff,et al.  Tumorigenesis and Neoplastic Progression Analysis of Orthologous Gene Expression between Human Pulmonary Adenocarcinoma and a Carcinogen-Induced Murine Model , 2010 .

[39]  K. Nakayama,et al.  Ubiquitin ligases: cell-cycle control and cancer , 2006, Nature Reviews Cancer.

[40]  Pang-Kuo Lo,et al.  Epigenetic suppression of secreted frizzled related protein 1 (SFRP1) expression in human breast cancer , 2006, Cancer biology & therapy.

[41]  Carlos Cordon-Cardo,et al.  Defining molecular profiles of poor outcome in patients with invasive bladder cancer using oligonucleotide microarrays. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[42]  Mahesh C Sharma,et al.  Roscovitine regulates invasive breast cancer cell (MDA-MB231) proliferation and survival through cell cycle regulatory protein cdk5. , 2007, Experimental and molecular pathology.

[43]  E. Lam,et al.  The emerging roles of forkhead box (Fox) proteins in cancer , 2007, Nature Reviews Cancer.

[44]  Alicia Zhou,et al.  Mesenchyme Forkhead 1 (FOXC2) plays a key role in metastasis and is associated with aggressive basal-like breast cancers , 2007, Proceedings of the National Academy of Sciences.

[45]  B. Burgering,et al.  Stressing the role of FoxO proteins in lifespan and disease , 2007, Nature Reviews Molecular Cell Biology.

[46]  A. Ashworth,et al.  BRCA1 dysfunction in sporadic basal-like breast cancer , 2007, Oncogene.

[47]  G. Sumara,et al.  A Cul3-based E3 ligase removes Aurora B from mitotic chromosomes, regulating mitotic progression and completion of cytokinesis in human cells. , 2007, Developmental cell.