PSA regulates androgen receptor expression in prostate cancer cells†‡§
Parmita Saxena
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, Massachusetts
Search for more papers by this authorMarco Trerotola
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania
Search for more papers by this authorTao Wang
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Department of Radiation Oncology, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorJing Li
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorAejaz Sayeed
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania
Search for more papers by this authorJennifer VanOudenhove
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorDave S. Adams
Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, Massachusetts
Search for more papers by this authorThomas J. FitzGerald
Department of Radiation Oncology, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorDario C. Altieri
Prostate Cancer Discovery and Development Program, The Wistar Institute Cancer Center, Philadelphia, Pennsylvania
Search for more papers by this authorCorresponding Author
Lucia R. Languino
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania
Department of Cancer Biology, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107.Search for more papers by this authorParmita Saxena
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, Massachusetts
Search for more papers by this authorMarco Trerotola
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania
Search for more papers by this authorTao Wang
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Department of Radiation Oncology, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorJing Li
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorAejaz Sayeed
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania
Search for more papers by this authorJennifer VanOudenhove
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorDave S. Adams
Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, Massachusetts
Search for more papers by this authorThomas J. FitzGerald
Department of Radiation Oncology, University of Massachusetts Medical School, Worcester, Massachusetts
Search for more papers by this authorDario C. Altieri
Prostate Cancer Discovery and Development Program, The Wistar Institute Cancer Center, Philadelphia, Pennsylvania
Search for more papers by this authorCorresponding Author
Lucia R. Languino
Department of Cancer Biology, Prostate Cancer Discovery and Development Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania
Department of Cancer Biology, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107.Search for more papers by this authorParmita Saxena and Marco Trerotola contributed equally to this work.
Disclosure: The authors declare no potential conflict of interest.
P. Saxena is currently employed by Novartis Vaccines and Diagnostics.
Abstract
BACKGROUND
Prostate-specific antigen (PSA) is a pivotal downstream target gene of the androgen receptor (AR), and a serum biomarker to monitor prostate cancer (PrCa) progression. It has been reported that PSA transactivates AR, but the mechanistic requirements of this response have not been investigated.
METHODS
We studied the localization of PSA, AR, and Src in intracellular compartments of synthetic androgen (R1881)-stimulated LNCaP and C4-2B PrCa cells, using immunofluorescence and subcellular fractionation approaches. We also investigated the effect of downregulation of PSA on AR expression by immunoblotting and real-time PCR using short hairpin RNA (shRNA) and small interfering RNA (siRNA). Src activity was analyzed by immunoblotting.
RESULTS
R1881 stimulation induced nuclear localization of both PSA and AR in LNCaP and C4-2B PrCa cells as well as increased phosphorylation of Src. Stable shRNA or transient siRNA knockdown of PSA resulted in reduced AR protein levels as well as AR mRNA levels in C4-2B cells. Similar to C4-2B cells, ablation of AR levels upon silencing of PSA was also confirmed in VCaP cells, another androgen-independent cell line. Silencing of PSA did not cause significant changes in Src activation; besides, Src regulation by integrins did not appear to affect AR transcriptional activity.
CONCLUSIONS
PSA localizes to nuclei of androgen-stimulated PrCa cells, and controls AR mRNA and protein levels. This regulatory loop is specific for PSA, does not involve known AR activators such as Src and AKT, and may contribute to AR signaling under conditions of increasing PSA levels in patients. Prostate 72:769–776, 2012. © 2011 Wiley Periodicals, Inc.
REFERENCES
- 1 Feldman BJ, Feldman D. The development of androgen-independent prostate cancer. Nat Rev Cancer 2001; 1(1): 34–45.
- 2 Heinlein CA, Chang C. Androgen receptor in prostate cancer. Endocr Rev 2004; 25(2): 276–308.
- 3 Williams SA, Singh P, Isaacs JT, Denmeade SR. Does PSA play a role as a promoting agent during the initiation and/or progression of prostate cancer? Prostate 2007; 67(3): 312–329.
- 4 Lawrence MG, Lai J, Clements JA. Kallikreins on steroids: Structure, function, and hormonal regulation of prostate-specific antigen and the extended kallikrein locus. Endocr Rev 2010; 31(4): 407–446.
- 5 Sotiropoulou G, Pampalakis G, Diamandis EP. Functional roles of human kallikrein-related peptidases. J Biol Chem 2009; 284(48): 32989–32994.
- 6 Kumar A, Mikolajczyk SD, Goel AS, Millar LS, Saedi MS. Expression of pro form of prostate-specific antigen by mammalian cells and its conversion to mature, active form by human kallikrein 2. Cancer Res 1997; 57(15): 3111–3114.
- 7 Lovgren J, Rajakoski K, Karp M, Lundwall A, Lilja H. Activation of the zymogen form of prostate-specific antigen by human glandular kallikrein 2. Biochem Biophys Res Commun 1997; 238(2): 549–555.
- 8 Kim J, Coetzee GA. Prostate specific antigen gene regulation by androgen receptor. J Cell Biochem 2004; 93(2): 233–241.
- 9 Yousef GM, Diamandis EP. The new human tissue kallikrein gene family: Structure, function, and association to disease. Endocr Rev 2001; 22(2): 184–204.
- 10 Nash AF, Melezinek I. The role of prostate specific antigen measurement in the detection and management of prostate cancer. Endocr Relat Cancer 2000; 7(1): 37–51.
- 11 Ryan CJ, Smith A, Lal P, Satagopan J, Reuter V, Scardino P, Gerald W, Scher HI. Persistent prostate-specific antigen expression after neoadjuvant androgen depletion: An early predictor of relapse or incomplete androgen suppression. Urology 2006; 68(4): 834–839.
- 12 Niu Y, Yeh S, Miyamoto H, Li G, Altuwaijri S, Yuan J, Han R, Ma T, Kuo HC, Chang C. Tissue prostate-specific antigen facilitates refractory prostate tumor progression via enhancing ARA70-regulated androgen receptor transactivation. Cancer Res 2008; 68(17): 7110–7119.
- 13 Fizazi K. The role of Src in prostate cancer. Ann Oncol 2007; 18(11): 1765–1773.
- 14 Lee LF, Louie MC, Desai SJ, Yang J, Chen HW, Evans CP, Kung HJ. Interleukin-8 confers androgen-independent growth and migration of LNCaP: Differential effects of tyrosine kinases Src and FAK. Oncogene 2004; 23(12): 2197–2205.
- 15 Guo Z, Dai B, Jiang T, Xu K, Xie Y, Kim O, Nesheiwat I, Kong X, Melamed J, Handratta VD, Njar VC, Brodie AM, Yu LR, Veenstra TD, Chen H, Qiu Y. Regulation of androgen receptor activity by tyrosine phosphorylation. Cancer Cell 2006; 10(4): 309–319.
- 16 Desgrosellier JS, Barnes LA, Shields DJ, Huang M, Lau SK, Prevost N, Tarin D, Shattil SJ, Cheresh DA. An integrin α(v)β(3)-c-Src oncogenic unit promotes anchorage-independence and tumor progression. Nat Med 2009; 15(10): 1163–1169.
- 17 Zheng DQ, Woodard AS, Fornaro M, Tallini G, Languino LR. Prostatic carcinoma cell migration via αvβ3 integrin is modulated by a focal adhesion kinase pathway. Cancer Res 1999; 59(7): 1655–1664.
- 18 Cole CN, Hammell CM. Nucleocytoplasmic transport: Driving and directing transport. Curr Biol 1998; 8(11): R368–R372.
- 19 Hao Y, Macara IG. Regulation of chromatin binding by a conformational switch in the tail of the Ran exchange factor RCC1. J Cell Biol 2008; 182(5): 827–836.
- 20 Korenchuk S, Lehr JE, MClean L, Lee YG, Whitney S, Vessella R, Lin DL, Pienta KJ. VCaP, a cell-based model system of human prostate cancer. In Vivo 2001; 15(2): 163–168.
- 21 Fornaro M, Manzotti M, Tallini G, Slear AE, Bosari S, Ruoslahti E, Languino LR. β1C integrin in epithelial cells correlates with a nonproliferative phenotype: Forced expression of β1C inhibits prostate epithelial cell proliferation. Am J Pathol 1998; 153(4): 1079–1087.
- 22 Goel HL, Fornaro M, Moro L, Teider N, Rhim JS, King M, Languino LR. Selective modulation of type 1 insulin-like growth factor receptor signaling and functions by β1 integrins. J Cell Biol 2004; 166(3): 407–418.
- 23 Sayeed A, Alam N, Trerotola M, Languino LR. Insulin-like growth factor 1 stimulation of androgen receptor activity requires β1A integrins. J Cell Physiol 2011; (in press).
- 24 Xi Z, Klokk TI, Korkmaz K, Kurys P, Elbi C, Risberg B, Danielsen H, Loda M, Saatcioglu F. Kallikrein 4 is a predominantly nuclear protein and is overexpressed in prostate cancer. Cancer Res 2004; 64(7): 2365–2370.
- 25 Linstedt AD, Foguet M, Renz M, Seelig HP, Glick BS, Hauri HP. A C-terminally-anchored Golgi protein is inserted into the endoplasmic reticulum and then transported to the Golgi apparatus. Proc Natl Acad Sci USA 1995; 92(11): 5102–5105.
- 26 Linstedt AD, Hauri HP. Giantin, a novel conserved Golgi membrane protein containing a cytoplasmic domain of at least 350kDa. Mol Biol Cell 1993; 4(7): 679–693.
- 27 Yang L, Xie S, Jamaluddin MS, Altuwaijri S, Ni J, Kim E, Chen YT, Hu YC, Wang L, Chuang KH, Wu CT, Chang C. Induction of androgen receptor expression by phosphatidylinositol 3-kinase/Akt downstream substrate, FOXO3a, and their roles in apoptosis of LNCaP prostate cancer cells. J Biol Chem 2005; 280(39): 33558–33565.
- 28 Watson PA, Chen YF, Balbas MD, Wongvipat J, Socci ND, Viale A, Kim K, Sawyers CL. Constitutively active androgen receptor splice variants expressed in castration-resistant prostate cancer require full-length androgen receptor. Proc Natl Acad Sci USA 2010; 107(39): 16759–16765.
- 29 Guo Z, Yang X, Sun F, Jiang R, Linn DE, Chen H, Kong X, Melamed J, Tepper CG, Kung HJ, Brodie AM, Edwards J, Qiu Y. A novel androgen receptor splice variant is up-regulated during prostate cancer progression and promotes androgen depletion-resistant growth. Cancer Res 2009; 69(6): 2305–2313.
- 30 Goel HL, Li J, Kogan S, Languino LR. Integrins in prostate cancer progression. Endocr Relat Cancer 2008; 15(3): 657–664.
- 31 Dong Y, Bui LT, Odorico DM, Tan OL, Myers SA, Samaratunga H, Gardiner RA, Clements JA. Compartmentalized expression of kallikrein 4 (KLK4/hK4) isoforms in prostate cancer: Nuclear, cytoplasmic and secreted forms. Endocr Relat Cancer 2005; 12(4): 875–889.
- 32 Williams SA, Jelinek CA, Litvinov I, Cotter RJ, Isaacs JT, Denmeade SR. Enzymatically active prostate-specific antigen promotes growth of human prostate cancers. Prostate 2011; (in press).
- 33 Sanda MG, Smith DC, Charles LG, Hwang C, Pienta KJ, Schlom J, Milenic D, Panicali D, Montie JE. Recombinant vaccinia-PSA (PROSTVAC) can induce a prostate-specific immune response in androgen-modulated human prostate cancer. Urology 1999; 53(2): 260–266.
- 34 Gulley J, Chen AP, Dahut W, Arlen PM, Bastian A, Steinberg SM, Tsang K, Panicali D, Poole D, Schlom J, Michael Hamilton J. Phase I study of a vaccine using recombinant vaccinia virus expressing PSA (rV-PSA) in patients with metastatic androgen-independent prostate cancer. Prostate 2002; 53(2): 109–117.
- 35 Lubaroff DM, Konety BR, Link B, Gerstbrein J, Madsen T, Shannon M, Howard J, Paisley J, Boeglin D, Ratliff TL, Williams RD. Phase I clinical trial of an adenovirus/prostate-specific antigen vaccine for prostate cancer: Safety and immunologic results. Clin Cancer Res 2009; 15(23): 7375–7380.
- 36 Kantoff PW, Schuetz TJ, Blumenstein BA, Glode LM, Bilhartz DL, Wyand M, Manson K, Panicali DL, Laus R, Schlom J, Dahut WL, Arlen PM, Gulley JL, Godfrey WR. Overall survival analysis of a phase II randomized controlled trial of a Poxviral-based PSA-targeted immunotherapy in metastatic castration-resistant prostate cancer. J Clin Oncol 2010; 28(7): 1099–1105.
- 37 Nesslinger NJ, Ng A, Tsang KY, Ferrara T, Schlom J, Gulley JL, Nelson BH. A viral vaccine encoding prostate-specific antigen induces antigen spreading to a common set of self-proteins in prostate cancer patients. Clin Cancer Res 2010; 16(15): 4046–4056.
- 38 Migliaccio A, Castoria G, Di Domenico M, de Falco A, Bilancio A, Lombardi M, Barone MV, Ametrano D, Zannini MS, Abbondanza C, Auricchio F. Steroid-induced androgen receptor-oestradiol receptor β-Src complex triggers prostate cancer cell proliferation. EMBO J 2000; 19(20): 5406–5417.
- 39 Lee LF, Guan J, Qiu Y, Kung HJ. Neuropeptide-induced androgen independence in prostate cancer cells: Roles of nonreceptor tyrosine kinases Etk/Bmx, Src, and focal adhesion kinase. Mol Cell Biol 2001; 21(24): 8385–8397.
- 40 Unni E, Sun S, Nan B, McPhaul MJ, Cheskis B, Mancini MA, Marcelli M. Changes in androgen receptor nongenotropic signaling correlate with transition of LNCaP cells to androgen independence. Cancer Res 2004; 64(19): 7156–7168.
