FOXA1 Promotes Tumor Progression in Prostate Cancer via the Insulin-Like Growth Factor Binding Protein 3 Pathway

Fork-head box protein A1 (FOXA1) is a “pioneer factor” that is known to bind to the androgen receptor (AR) and regulate the transcription of AR-specific genes. However, the precise role of FOXA1 in prostate cancer (PC) remains unknown. In this study, we report that FOXA1 plays a critical role in PC cell proliferation. The expression of FOXA1 was higher in PC than in normal prostate tissues (P = 0.0002), and, using immunohistochemical analysis, we found that FOXA1 was localized in the nucleus. FOXA1 expression levels were significantly correlated with both PSA and Gleason scores (P = 0.016 and P = 0.031, respectively). Moreover, FOXA1 up-regulation was a significant factor in PSA failure (P = 0.011). Depletion of FOXA1 in a prostate cancer cell line (LNCaP) using small interfering RNA (siRNA) significantly inhibited AR activity, led to cell-growth suppression, and induced G0/G1 arrest. The anti-proliferative effect of FOXA1 siRNA was mediated through insulin-like growth factor binding protein 3 (IGFBP-3). An increase in IGFBP-3, mediated by depletion of FOXA1, inhibited phosphorylation of MAPK and Akt, and increased expression of the cell cycle regulators p21 and p27. We also found that the anti-proliferative effect of FOXA1 depletion was significantly reversed by simultaneous siRNA depletion of IGFBP-3. These findings provide direct physiological and molecular evidence for a role of FOXA1 in controlling cell proliferation through the regulation of IGFBP-3 expression in PC.

[1]  K. Zaret,et al.  Specific interactions of the wing domains of FOXA1 transcription factor with DNA. , 2007, Journal of molecular biology.

[2]  Clifford A. Meyer,et al.  Androgen Receptor Regulates a Distinct Transcription Program in Androgen-Independent Prostate Cancer , 2009, Cell.

[3]  Nicholas Bruchovsky,et al.  Ligand-independent Activation of the Androgen Receptor by Interleukin-6 and the Role of Steroid Receptor Coactivator-1 in Prostate Cancer Cells* , 2002, The Journal of Biological Chemistry.

[4]  P. Nelson,et al.  Interaction of IGF signaling and the androgen receptor in prostate cancer progression , 2006, Journal of cellular biochemistry.

[5]  M. Gadelha,et al.  IGF-I, insulin and prostate cancer. , 2009, Arquivos brasileiros de endocrinologia e metabologia.

[6]  K. Uzawa,et al.  Characteristic gene expression profiles of benign prostatic hypertrophy and prostate cancer. , 2009, International journal of oncology.

[7]  T. Ichikawa,et al.  Hormone treatment for prostate cancer: current issues and future directions , 2005, Cancer Chemotherapy and Pharmacology.

[8]  J. Clifford,et al.  Direct Functional Interactions between Insulin-like Growth Factor-binding Protein-3 and Retinoid X Receptor-α Regulate Transcriptional Signaling and Apoptosis* , 2000, The Journal of Biological Chemistry.

[9]  R. Caprioli,et al.  Forkhead box A1 regulates prostate ductal morphogenesis and promotes epithelial cell maturation , 2005, Development.

[10]  J. Mirosevich,et al.  Expression of Foxa transcription factors in the developing and adult murine prostate , 2005, The Prostate.

[11]  R. Matusik,et al.  Upstream stimulatory factor 2, a novel FoxA1-interacting protein, is involved in prostate-specific gene expression. , 2009, Molecular endocrinology.

[12]  Glen Kristiansen,et al.  Tumorigenesis and Neoplastic Progression FOXA 1 Promotes Tumor Progression in Prostate Cancer and Represents a Novel Hallmark of Castration-Resistant Prostate Cancer , 2012 .

[13]  T. Hickey,et al.  FOXA1: master of steroid receptor function in cancer , 2011, The EMBO journal.

[14]  P. Cohen,et al.  Insulin-like Growth Factor (IGF)-binding Protein-3 Induces Apoptosis and Mediates the Effects of Transforming Growth Factor-β1 on Programmed Cell Death through a p53- and IGF-independent Mechanism* , 1997, The Journal of Biological Chemistry.

[15]  M. Gleave,et al.  Derivation of androgen‐independent human LNCaP prostatic cancer cell sublines: Role of bone stromal cells , 1994, International journal of cancer.

[16]  David E. Misek,et al.  The hepatocyte nuclear factor 3 alpha gene, HNF3alpha (FOXA1), on chromosome band 14q13 is amplified and overexpressed in esophageal and lung adenocarcinomas. , 2002, Cancer research.

[17]  D. Feldman,et al.  The development of androgen-independent prostate cancer , 2001, Nature Reviews Cancer.

[18]  A. Gao,et al.  Molecular mechanisms of castration-resistant prostate cancer progression. , 2009, Future oncology.

[19]  Hong Xin,et al.  DU‐145 and PC‐3 human prostate cancer cell lines express androgen receptor: Implications for the androgen receptor functions and regulation , 2006, FEBS letters.

[20]  Rajiv Dhir,et al.  Talin1 promotes tumor invasion and metastasis via focal adhesion signaling and anoikis resistance. , 2010, Cancer research.

[21]  R. Baxter,et al.  Cellular actions of the insulin-like growth factor binding proteins. , 2002, Endocrine reviews.

[22]  O. Kallioniemi,et al.  Dual role of FoxA1 in androgen receptor binding to chromatin, androgen signalling and prostate cancer , 2011, The EMBO journal.

[23]  E. Keller,et al.  Prostate cancer stromal cells and LNCaP cells coordinately activate the androgen receptor through synthesis of testosterone and dihydrotestosterone from dehydroepiandrosterone. , 2009, Endocrine-related cancer.

[24]  J. Reis-Filho,et al.  Forkhead box A1 expression in breast cancer is associated with luminal subtype and good prognosis , 2007, Journal of Clinical Pathology.

[25]  M. Webber,et al.  Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. , 1997, Carcinogenesis.

[26]  M. Gleave,et al.  Differential regulation of IGFBP‐3 by the androgen receptor in the lineage‐related androgen‐dependent LNCaP and androgen‐independent C4‐2 prostate cancer models , 2006, The Prostate.

[27]  J. Silha,et al.  Insulin-like growth factor (IGF) binding protein-3 attenuates prostate tumor growth by IGF-dependent and IGF-independent mechanisms. , 2006, Endocrinology.

[28]  G. Murphy,et al.  LNCaP model of human prostatic carcinoma. , 1983, Cancer research.

[29]  Brad T. Sherman,et al.  Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists , 2008, Nucleic acids research.

[30]  Thomas Wheeler,et al.  High Level of Androgen Receptor Is Associated With Aggressive Clinicopathologic Features and Decreased Biochemical Recurrence-free Survival in Prostate: Cancer Patients Treated With Radical Prostatectomy , 2004, The American journal of surgical pathology.

[31]  P. Yamada,et al.  Perspectives in mammalian IGFBP-3 biology: local vs. systemic action. , 2009, American journal of physiology. Cell physiology.

[32]  Wei Li,et al.  Definition of a FoxA1 Cistrome that is crucial for G1 to S-phase cell-cycle transit in castration-resistant prostate cancer. , 2011, Cancer research.

[33]  M. Loda,et al.  FOXA1 Is a Potential Oncogene in Anaplastic Thyroid Carcinoma , 2009, Clinical Cancer Research.

[34]  L. Tran,et al.  Cell autonomous role of PTEN in regulating castration-resistant prostate cancer growth. , 2011, Cancer cell.

[35]  M. Lupien,et al.  Cistromics of hormone-dependent cancer. , 2009, Endocrine-related cancer.

[36]  Brad T. Sherman,et al.  Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources , 2008, Nature Protocols.

[37]  K. Uzawa,et al.  State of homeobox A10 expression as a putative prognostic marker for oral squamous cell carcinoma. , 2009, Oncology reports.

[38]  David E. Misek,et al.  The Hepatocyte Nuclear Factor 3 α Gene, HNF3α (FOXA1), on Chromosome Band 14q13 Is Amplified and Overexpressed in Esophageal and Lung Adenocarcinomas , 2002 .

[39]  Y. Furuya,et al.  Implications of insulin‐like growth factor‐I for prostate cancer therapies , 2009, International journal of urology : official journal of the Japanese Urological Association.

[40]  D. Reich,et al.  Functional Enhancers at the Gene-Poor 8q24 Cancer-Linked Locus , 2009, PLoS genetics.

[41]  S. Badve,et al.  High‐level expression of forkhead‐box protein A1 in metastatic prostate cancer , 2011, Histopathology.

[42]  J. Mirosevich,et al.  The Role of Foxa Proteins in the Regulation of Androgen Receptor Activity , 2009 .

[43]  T. Ishigami,et al.  Epithelial Cell Transforming Sequence 2 in Human Oral Cancer , 2010, PloS one.

[44]  Renjie Jin,et al.  The role of hepatocyte nuclear factor-3 alpha (Forkhead Box A1) and androgen receptor in transcriptional regulation of prostatic genes. , 2003, Molecular endocrinology.

[45]  N. Kyprianou,et al.  Androgen receptor and growth factor signaling cross-talk in prostate cancer cells. , 2008, Endocrine-related cancer.

[46]  Charles M Perou,et al.  FOXA1 Expression in Breast Cancer—Correlation with Luminal Subtype A and Survival , 2007, Clinical Cancer Research.

[47]  Guido Jenster,et al.  Gene expression of forkhead transcription factors in the normal and diseased human prostate , 2009, BJU international.