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.
References
[1]
Mulholland DJ, Tran LM, Li Y, Cai H, Morim A, et al. (2011) Cell autonomous role of PTEN in regulating castration-resistant prostate cancer growth. Cancer Cell 19: 792–804.
[2]
Li R, Wheeler T, Dai H, Frolov A, Thompson T, et al. (2004) 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. Am J Surg Pathol 28: 928–934.
[3]
Dutt SS, Gao AC (2009) Molecular mechanisms of castration-resistant prostate cancer progression. Future Oncol 5: 1403–1413.
[4]
Endo T, Uzawa K, Suzuki H, Tanzawa H, Ichikawa T (2009) Characteristic gene expression profiles of benign prostatic hypertrophy and prostate cancer. Int J Oncol 35: 499–509.
[5]
Augello MA, Hickey TE, Knudsen KE (2011) FOXA1: master of steroid receptor function in cancer. EMBO J 30: 3885–3894.
[6]
Gao N, Zhang J, Rao MA, Case TC, Mirosevich J, et al. (2003) The role of hepatocyte nuclear factor-3 alpha (Forkhead Box A1) and androgen receptor in transcriptional regulation of prostatic genes. Mol Endocrinol 17: 1484–1507.
[7]
Lupien M, Brown M (2009) Cistromics of hormone-dependent cancer. Endocr Relat Cancer 16: 381–389.
[8]
Cirillo LA, Zaret KS (2007) Specific interactions of the wing domains of FOXA1 transcription factor with DNA. J Mol Biol 366: 720–724.
[9]
Degraff DJ, Yu X, Sun Q, Mirosevich J, Jin JR, et al. (2009) The Role of Foxa Proteins in the Regulation of Androgen Receptor Activity. Androgen Action in Prostate Cancer Part 6: 587–615.
[10]
Gao N, Ishii K, Mirosevich J, Kuwajima S, Oppenheimer SR, et al. (2005) Forkhead box A1 regulates prostate ductal morphogenesis and promotes epithelial cell maturation. Development 132: 3431–3443.
[11]
Badve S, Turbin D, Thorat MA, Morimiya A, Nielsen TO, et al. (2007) FOXA1 expression in breast cancer–correlation with luminal subtype A and survival. Clin Cancer Res 13: 4415–4421.
[12]
Thorat MA, Marchio C, Morimiya A, Savage K, Nakshatri H, et al. (2008) Forkhead box A1 expression in breast cancer is associated with luminal subtype and good prognosis. J Clin Pathol 61: 327–332.
[13]
Lin L, Miller CT, Contreras JI, Prescott MS, Dagenais SL, et al. (2002) The hepatocyte nuclear factor 3 alpha gene, HNF3alpha (FOXA1), on chromosome band 14q13 is amplified and overexpressed in esophageal and lung adenocarcinomas. Cancer Res 62: 5273–5279.
[14]
Nucera C, Eeckhoute J, Finn S, Carroll JS, Ligon AH, et al. (2009) FOXA1 is a potential oncogene in anaplastic thyroid carcinoma. Clin Cancer Res 15: 3680–3689.
[15]
Gerhardt J, Montani M, Wild P, Beer M, Huber F, et al. (2012) FOXA1 promotes tumor progression in prostate cancer and represents a novel hallmark of castration-resistant prostate cancer. Am J Pathol 180: 848–861.
[16]
Horoszewicz JS, Leong SS, Kawinski E, Karr JP, Rosenthal H, et al. (1983) LNCaP model of human prostatic carcinoma. Cancer Res 43: 1809–1818.
[17]
Wu HC, Hsieh JT, Gleave ME, Brown NM, Pathak S, et al. (1994) Derivation of androgen-independent human LNCaP prostatic cancer cell sublines: role of bone stromal cells. Int J Cancer 57: 406–412.
[18]
Alimirah F, Chen J, Basrawala Z, Xin H, Choubey D (2006) DU-145 and PC-3 human prostate cancer cell lines express androgen receptor: implications for the androgen receptor functions and regulation. FEBS Lett 580: 2294–2300.
[19]
Bello D, Webber MM, Kleinman HK, Wartinger DD, Rhim JS (1997) Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. Carcinogenesis 18: 1215–1223.
[20]
Silha JV, Sheppard PC, Mishra S, Gui Y, Schwartz J, et al. (2006) Insulin-like growth factor (IGF) binding protein-3 attenuates prostate tumor growth by IGF-dependent and IGF-independent mechanisms. Endocrinology 147: 2112–2121.
[21]
Kojima S, Inahara M, Suzuki H, Ichikawa T, Furuya Y (2009) Implications of insulin-like growth factor-I for prostate cancer therapies. Int J Urol 16: 161–167.
[22]
Lima GA, Correa LL, Gabrich R, Miranda LC, Gadelha MR (2009) IGF-I, insulin and prostate cancer. Arq Bras Endocrinol Metabol 53: 969–975.
[23]
Firth SM, Baxter RC (2002) Cellular actions of the insulin-like growth factor binding proteins. Endocr Rev 23: 824–854.
[24]
Kojima S, Mulholland DJ, Ettinger S, Fazli L, Nelson CC, et al. (2006) Differential regulation of IGFBP-3 by the androgen receptor in the lineage-related androgen-dependent LNCaP and androgen-independent C4-2 prostate cancer models. Prostate 66: 971–986.
[25]
Jia L, Landan G, Pomerantz M, Jaschek R, Herman P, et al. (2009) Functional enhancers at the gene-poor 8q24 cancer-linked locus. PLoS Genet 5: e1000597.
[26]
Sun Q, Yu X, Degraff DJ, Matusik RJ (2009) Upstream stimulatory factor 2, a novel FoxA1-interacting protein, is involved in prostate-specific gene expression. Mol Endocrinol 23: 2038–2047.
[27]
Mirosevich J, Gao N, Matusik RJ (2005) Expression of Foxa transcription factors in the developing and adult murine prostate. Prostate 62: 339–352.
[28]
Sahu B, Laakso M, Ovaska K, Mirtti T, Lundin J, et al. (2011) Dual role of FoxA1 in androgen receptor binding to chromatin, androgen signalling and prostate cancer. EMBO J 30: 3962–3976.
[29]
Jain RK, Mehta RJ, Nakshatri H, Idrees MT, Badve SS (2011) High-level expression of forkhead-box protein A1 in metastatic prostate cancer. Histopathology 58: 766–772.
[30]
van der Heul-Nieuwenhuijsen L, Dits NF, Jenster G (2009) Gene expression of forkhead transcription factors in the normal and diseased human prostate. BJU Int 103: 1574–1580.
[31]
Wang Q, Li W, Zhang Y, Yuan X, Xu K, et al. (2009) Androgen receptor regulates a distinct transcription program in androgen-independent prostate cancer. Cell 138: 245–256.
[32]
Yamada PM, Lee KW (2009) Perspectives in mammalian IGFBP-3 biology: local vs. systemic action. Am J Physiol Cell Physiol 296: C954–976.
[33]
Rajah R, Valentinis B, Cohen P (1997) Insulin-like growth factor (IGF)-binding protein-3 induces apoptosis and mediates the effects of transforming growth factor-beta1 on programmed cell death through a p53- and IGF-independent mechanism. J Biol Chem 272: 12181–12188.
[34]
Liu B, Lee HY, Weinzimer SA, Powell DR, Clifford JL, et al. (2000) Direct functional interactions between insulin-like growth factor-binding protein-3 and retinoid X receptor-alpha regulate transcriptional signaling and apoptosis. J Biol Chem 275: 33607–33613.
[35]
Feldman BJ, Feldman D (2001) The development of androgen-independent prostate cancer. Nat Rev Cancer 1: 34–45.
[36]
Ueda T, Mawji NR, Bruchovsky N, Sadar MD (2002) Ligand-independent activation of the androgen receptor by interleukin-6 and the role of steroid receptor coactivator-1 in prostate cancer cells. J Biol Chem 277: 38087–38094.
[37]
Zhu ML, Kyprianou N (2008) Androgen receptor and growth factor signaling cross-talk in prostate cancer cells. Endocr Relat Cancer 15: 841–849.
[38]
Wu JD, Haugk K, Woodke L, Nelson P, Coleman I, et al. (2006) Interaction of IGF signaling and the androgen receptor in prostate cancer progression. J Cell Biochem 99: 392–401.
[39]
Zhang C, Wang L, Wu D, Chen H, Chen Z, et al. (2011) Definition of a FoxA1 Cistrome that is crucial for G1 to S-phase cell-cycle transit in castration-resistant prostate cancer. Cancer Res 71: 6738–6748.
[40]
Ichikawa T, Suzuki H, Ueda T, Komiya A, Imamoto T, et al. (2005) Hormone treatment for prostate cancer: current issues and future directions. Cancer Chemother Pharmacol 56 Suppl 158–63.
[41]
Mizokami A, Koh E, Izumi K, Narimoto K, Takeda M, et al. (2009) Prostate cancer stromal cells and LNCaP cells coordinately activate the androgen receptor through synthesis of testosterone and dihydrotestosterone from dehydroepiandrosterone. Endocr Relat Cancer 16: 1139–1155.
[42]
Sakamoto S, McCann RO, Dhir R, Kyprianou N (2010) Talin1 promotes tumor invasion and metastasis via focal adhesion signaling and anoikis resistance. Cancer Res 70: 1885–1895.
[43]
Iyoda M, Kasamatsu A, Ishigami T, Nakashima D, Endo-Sakamoto Y, et al. (2010) Epithelial cell transforming sequence 2 in human oral cancer. PLoS One 5: e14082.
[44]
Yamatoji M, Kasamatsu A, Yamano Y, Sakuma K, Ogoshi K, et al. (2010) State of homeobox A10 expression as a putative prognostic marker for oral squamous cell carcinoma. Oncol Rep 23: 61–67.
[45]
Huang da W, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4: 44–57.
[46]
Huang da W, Sherman BT, Lempicki RA (2009) Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res 37: 1–13.
