[en] Androgens control both growth and differentiation of the normal prostate gland. However, the mechanisms by which androgens act upon the cell cycle machinery to regulate these two fundamental processes are largely unknown. The cyclin-dependent kinase (cdk) inhibitor p27 is a negative cell cycle regulator involved in differentiation-associated growth arrest. Here, we investigate the role and regulation of p27 in the testosterone proprionate (TP)-stimulated regeneration of the ventral prostate (VP) of castrated rats. Continuous TP administration to castrated rats triggered epithelial cell proliferation, which peaked at 72 h, and then declined despite further treatment. Castration-induced atrophy of the VP was associated with a significant increase in p27 expression as compared with the VP of intact animals. Twelve hours after the initiation of androgen treatment, total p27 levels as well as its fraction bound to cdk2, its main target, significantly dropped in the VP of castrated rats. Thereafter, concomitantly to the induction of epithelial cell proliferation, the glandular morphology of VP was progressively restored at 48-96 h of TP treatment. During this period of the regenerative process, whereas both proliferating basal and secretory epithelial cells did not express p27, the protein was selectively up-regulated in the nonproliferating secretory epithelial compartment. This up-regulation of p27 expression was coincident with an increase in its association with, and presumably inhibition of, cdk2. At each time point of TP treatment, p27 abundance in the VP was inversely correlated with the level of its proteasome-dependent degradation activity measured in vitro in VP lysates, whereas only slight changes in the amount of p27 transcripts were detected. In addition, the antiandrogen flutamide blocked maximal TP-induced p27 degradation completely. Finally, the expression of skp2, the ubiquitin ligase that targets p27 for degradation, was seen to increase with androgen administration, preceding maximal proliferation and concomitantly to augmented p27 degradation activity. Taken together, our data indicate that androgens mediate both proliferation and differentiation signals in normal prostate epithelial cells in vivo, through regulation of p27.
Disciplines :
Urology & nephrology
Author, co-author :
Waltregny, David ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Labo de recherche sur les métastases - Département des sciences biomédicales et précliniques
Leav, Irwin
Signoretti, Sabina
Soung, Peggy
Lin, Douglas
Merk, Frederick
Adams, Jason Y.
Bhattacharya, Nandita
Cirenei, Nicola
Loda, Massimo
Language :
English
Title :
Androgen-driven prostate epithelial cell proliferation and differentiation in vivo involve the regulation of p27.
Publication date :
2001
Journal title :
Molecular Endocrinology
ISSN :
0888-8809
eISSN :
1944-9917
Publisher :
Endocrine Society, Chevy Chase, United States - Maryland
Cunha G.R., Donjacour A.A., Cooke P.S., Mee S., Bigsby R.M., Higgins S.J., Sugimura Y. (1987) The endocrinology and developmental biology of the prostate. Endocr Rev 8:338-362.
Evans G.S., Chandler J.A. (1987) Cell proliferation studies in the rat prostate. I. The proliferative role of basal and secretory cells during normal growth. Prostate 10:163-178.
English H.F., Santen R.J., Isaacs J.T. (1987) Response of glandular versus basal rat ventral prostatic epithelial cells to androgen withdrawal and replacement. Prostate 11:229-242.
Isaacs J.T., Lundmo P.I., Berges R., Martikainen P., Kyprianou N., English H.F. (1992) Androgen regulation of programmed death of normal and malignant prostatic cells. J Androl 13:457-464.
Bruchovsky N., Lesser B., Van Dorne E., Craven S. (1975) Hormonal effects on cell proliferation in rat prostate. Vitam Horm 33:61-102.
Ho S.M., Damassa D., Kwan P.W., Seto H.S., Leav I. (1985) Androgen receptor levels and androgen contents in the prostate lobes of intact and testosterone-treated Noble rats. J Androl 6:279-290.
Chen Y., Robles A.I., Martinez L.A., Liu F., Gimenez-Conti I.B., Conti C.J. (1996) Expression of G1 cyclins, cyclin-dependent kinases, and cyclin-dependent kinase inhibitors in androgen-induced prostate proliferation in castrated rats. Cell Growth Differ 7:1571-1578.
Leav I., Merk F.B., Kwan P.W., Ho S.M. (1989) Androgen-supported estrogen-enhanced epithelial proliferation in the prostates of intact Noble rats. Prostate 15:23-40.
Wang X., Hsieh J.T. (1994) Androgen repression of cytokeratin gene expression during rat prostate differentiation: Evidence for an epithelial stem cell-associated marker. Chin Med Sci J 9:237-241.
English H.F., Drago J.R., Santen R.J. (1985) Cellular response to androgen depletion and repletion in the rat ventral prostate: Autoradiography and morphometric analysis. Prostate 7:41-51.
Katz A.E., Benson M.C., Wise G.J., Olsson C.A., Bandyk M.G., Sawczuk I.S., Tomashefsky P., Buttyan R. (1989) Gene activity during the early phase of androgen-stimulated rat prostate regrowth. Cancer Res 49:5889-5894.
Epner D.E., Partin A.W., Schalken J.A., Isaacs J.T., Coffey D.S. (1993) Association of glyceraldehyde-3-phosphate dehydrogenase expression with cell motility and metastatic potential of rat prostatic adenocarcinoma. Cancer Res 53:1995-1997.
Morgan D.O. (1997) Cyclin-dependent kinases: Engines, clocks, and microprocessors. Annu Rev Dev Biol 13:261-291.
Sherr C.J. (1996) Cancer cell cycles. Science 274:1672-1677.
DelSal G., Loda M., Pagano M. (1996) Cell cycle and cancer: Critical events at the G1 restriction point. Crit Rev Oncog 7:127-142.
Sherr C.J., Roberts J.M. (1999) CDK inhibitors: Positive and negative regulators of G1-phase progression. Genes Dev 13:1501-1512.
Hengst L., Reed S.I. (1998) Inhibitors of the Cip/Kip family. Curr Top Microbiol Immunol 227:25-41.
Polyak K., Kato J.Y., Solomon M.J., Sherr C.J., Massague J., Roberts J.M., Koff A. (1994) p27 a cyclin- Cdk inhibitor, links transforming growth factor-β and contact inhibition to cell cycle arrest. Genes Dev 8:9-22.
Polyak K., Lee M.-H., Erdjument-Bromage A., Koff A., Roberts J.M., Tempst P., Massague J. (1994) Cloning of p27, a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals. Cell 78:59-66.
Toyoshima H., Hunter T. (1994) p27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21. Cell 78:67-74.
Fang F., Orend G., Watanabe N., Hunter T., Ruoslahti E. (1996) Dependence of cyclin E-CDK2 kinase activity on cell anchorage. Science 271:499-502.
Kato J., Matsuoka M., Polyak K., Massague J., Sherr C.J. (1994) Cyclic AMP-induced G1 phase arrest mediated by an inhibitor (p27) of cyclin-dependent kinase 4 activation. Cell 79:487-496.
Kuniyasu H., Yasui W., Kitahara K., Naka K., Yokozaki H., Akama Y., Hamamoto T., Tahara H., Tahara E. (1997) Growth inhibitory effect of interferon-β is associated with the induction of cyclin-dependent kinase inhibitor p27 in a hyman gastric carcinoma cell line. Cell Growth Differ 8:47-52.
Harvat B.L., Seth P., Jetten A.M. (1997) The role of p27 in γ-interferon-mediated growth arrest of mammary epithelial cells and related defects in mammary carcinoma cells. Oncogene 14:2111-2122.
Kortylewski M., Heinrich P.C., Mackiewicz A., Schniertshauer U., Klingmuller U., Nakajima K., Hirano T., Horn F., Behrmann I. (1999) Interleukin-6 and oncostatin M-induced growth inhibition of human A375 melanoma cells is STAT-dependent and involves upregulation of the cyclin-dependent kinase inhibitor p27/KIP1. Oncogene 18:3742-3753.
Nourse J., Firpo E., Flanagan W.M., Coats S., Polyak K., Lee M.H., Massague J., Crabtree G.R., Roberts J.M. (1994) Interleukin-2-mediated elimination of the p27 cyclin-dependent kinase inhibitor prevented by rapamycin. Nature 372:570-573.
Coats S., Flanagan W.M., Nourse J., Roberts J.M. (1996) Requirement of p27 for restriction point control of the fibroblast cell cycle. Science 272:877-880.
Rivard N., L'Alleimain G., Bartek J., Pouyssegur J. (1996) Abrogation of p27 by cDNA antisense suppresses quiescence (G0 state) in fibroblasts. J Biol Chem 271:18337-18341.
Sheaff R.J., Groudine M., Gordon M., Roberts J.M., Clurman B.E. (1997) Cyclin E-CDK2 is a regulator of p27. Genes Dev 11:1464-1478.
Montagnoli A., Fiore F., Eytan E., Carrano A.C., Draetta G.F., Hershko A., Pagano M. (1999) Ubiquitination of p27 is regulated by Cdk-dependent phosphorylation and trimeric complex formation. Genes Dev 13:1181-1189.
Nguyen H., Gitig D.M., Koff A. (1999) Cell-free degradation of p27(kip1), a G1 cyclin-dependent kinase inhibitor, is dependent on CDK2 activity and the proteasome. Mol Cell Biol 19:1190-1201.
Vlach J., Hennecke S., Amati B. (1997) Phosphorylation-dependent degradation of the cyclin-dependent kinase inhibitor p27. EMBO J 16:5334-5344.
Carrano A.C., Eytan E., Hershko A., Pagano M. (1999) SKP2 is required for ubiquitin-mediated degradation of the CDK inhibitor p27. Nat Cell Biol 1:193-199.
Tsvetkov L.M., Yeh K.H., Lee S.J., Sun H., Zhang H. (1999) p27(Kip1) ubiquitination and degradation is regulated by the SCF(Skp2) complex through phosphorylated Thr187 in p27. Curr Biol 9:661-664.
Pagano M., Tam S.W., Theodoras A.M., Beer-Romero P., Del Sal G., Chau V., Yew P.R., Draetta G.F., Rolfe M. (1995) Role of the ubiquitin-proteasome pathway in regulating abundance of the cyclin-dependent kinase inhibitor p27. Science 269:682-685.
Medema R.H., Kops G.J., Bos J.L., Burgering B.M. (2000) AFX-Iike Forkhead transcription factors mediate cell-cycle regulation by Ras and PKB through p27. Nature 404:782-787.
Menjo M., Kaneko Y., Ogata E., Ikeda K., Nakanishi M. (1998) Critical role for p27 in cell cycle arrest after androgen depletion in mouse mammary carcinoma cells (SC-3). Oncogene 17:2619-2627.
Cordon-Cardo C., Koff A., Drobnjak M., Capodieci P., Osman I., Millard S.S., Gaudin P.B., Fazzari M., Zhang Z.F., Massague J., Scher H.I. (1998) Distinct altered patterns of p27 gene expression in benign prostatic hyperplasia and prostatic carcinoma. J Natl Cancer Inst 90:1284-1291.
Millard S.S., Yan J.S., Nguyen H., Pagano M., Kiyokawa H., Koff A. (1997) Enhanced ribosomal association of p27(Kip1) mRNA is a mechanism contributing to accumulation during growth arrest. J Biol Chem 272:7093-7098.
Hengst L., Reed S.I. (1996) Translational control of p27 accumulation during the cell cycle. Science 271:1861-1864.
Fero M.L., Rivkin M., Tasch M., Porter P., Carow C.E., Firpo E., Polyak K., Tsai L.H., Broudy V., Perlmutter R.M., Kaushansky K., Roberts J.M. (1996) A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(Kip1)-deficient mice. Cell 85:733-744.
Kiyokawa H., Kineman R.D., Manova-Todorova K.O., Soares V.C., Hoffman E.S., Ono M., Khanam D., Hayday A.C., Frohman L.A., Koff A. (1996) Enhanced growth of mice lacking the cyclin-dependent kinase inhibitor function of p27(Kip1). Cell 85:721-732.
Nakayama K., Ishida N., Shirane M., Inomata A., Inoue T., Shishido N., Horii I., Loh D.Y. (1996) Mice lacking p27(Kip1) display increased body size, multiple organ hyperplasia, retinal dysplasia, and pituitary tumors. Cell 85:707-720.
Tong W., Kiyokawa H., Soots T.J., Park M.S., Soares V.C., Manova K., Pollard J.W., Koff A. (1998) The absence of p27, an inhibitor of G1 cyclin-dependent kinases, uncouples differentiation and growth arrest during the granulosa-luteal transition. Cell Growth Differ 9:787-794.
Robker R.L., Richards J.S. (1998) Hormone-induced proliferation and differentiation of granulosa cells: A coordinated balance of the cell cycle regulators cyclin D2 and p27. Mol Endocrinol 12:924-940.
Casaccia-Bonnefil P., Tikoo R., Kiyokawa H., Friedrich V., Chao M.V., Koff A. (1997) Oligodendrocyte precursor differentiation is perturbed in the absence of the cyclin-dependent kinase inhibitor p27. Genes Dev 11:2335-2346.
Drissi H., Hushka D., Aslam F., Nguyen Q., Buffone E., Koff A., Van Wijnen A., Lian J.B., Stein J.L., Stein G.S. (1999) The cell cycle regulator p27 contributes to growth and differentiation of osteoblasts. Cancer Res 59:3705-3711.
Sgambato A., Cittadini A., Faraglia B., Weinstein I.B. (2000) Multiple functions of p27(Kip1) and its alterations in tumor cells: A review. J Cell Physiol 183:18-27.
Mori S., Murakami-Mori K., Bonavida B. (1999) Interleukin-6 induces G1 arrest through induction of p27(Kip1), a cyclin-dependent kinase inhibitor, and neuron-like morphology in LNCaP prostate tumor cells. Biochem Biophys Res Commun 257:609-614.
Zi X., Agarwal R. (1999) Silibinin decreases prostate-specific antigen with cell growth inhibition via G1 arrest, leading to differentiation of prostate carcinoma cells: Implications for prostate cancer intervention. Proc Natl Acad Sci USA 96:7490-7495.
De Marzo A.M., Meeker A.K., Epstein J.I., Coffey D.S. (1998) Prostate stem cell compartments: Expression of the cell cycle inhibitor p27 in normal, hyperplastic, and neoplastic cells. Am J Pathol 153:911-919.
Tsihlias J., Zhang W., Bhattacharya N., Flanagan M., Klotz L., Slingerland J. (2000) Involvement of p27 in G1 arrest by high dose 5 α-dihydrotestosterone in LNCaP human prostate cancer cells. Oncogene 19:670-679.
Kokontis J.M., Hay N., Liao S. (1998) Progression of LNCaP prostate tumor cells during androgen deprivation: Hormone-independent growth, repression of proliferation by androgen, and role for p27 in androgen-induced cell cycle arrest. Mol Endocrinol 12:941-953.
Agus D.B., Cordon-Cardo C., Fox W., Drobnjak M., Koff A., Golde D.W., Scher H.I. (1999) Prostate cancer cell cycle regulators: Response to androgen withdrawal and development of androgen independence. J Natl Cancer Inst 91:1869-1876.
Johnson M.A., Hernandez I., Wei Y., Greenberg N. (2000) Isolation and characterization of mouse probasin: An androgen-regulated protein specifically expressed in the differentiated prostate. Prostate 43:255-262.
Loda M., Cukor B., Tam S.W., Lavin P., Florentino M., Draetta G.F., Jessup J.M., Pagano M. (1997) Increased proteasome-dependent degradation of the cyclin-dependent kinase inhibitor p27 in aggressive colorectal carcinomas. Nat Med 3:231-234.
Maki C.G., Howley P.M. (1997) Ubiquitination of p53 and p21 is differentially affected by ionizing and UV radiation. Mol Cell Biol 17:355-363.
Maki C.G., Huibregtse J.M., Howley P.M. (1996) In vivo ubiquitination and proteasome-mediated degradation of p53. Cancer Res 56:2649-2654.
Yu Z.K., Gervais J.L., Zhang H. (1998) Human CUL-1 associates with the SKP1/SKP2 complex and regulates p21(CIP1/WAF1) and cyclin D proteins. Proc Natl Acad Sci USA 95:11324-11329.
Blagosklonny M.V., Wu G.S., Omura S., El-Deiry W.S. (1996) Proteosome-dependent regulation of p21/CIP1 expression. Biochem Biophys Res Commun 227:564-569.
Chung L.W.K., Coffey D.S. (1971) Biochemical characterization of prostatic nuclei. II. Relationship between DNA synthesis and protein synthesis. Biochim Biophys Acta 247:584.
Furuya Y., Lin X.S., Walsh J.C., Nelson W.G., Isaacs J.T. (1995) Androgen ablation-induced programmed death of prostatic glandular cells does not involve recruitment into a defective cell cycle or p53 induction. Endocrinology 136:1898-1906.
Evans G.S., Chandler J.A. (1987) Cell proliferation studies in the rat prostate. II. The effects of castration and androgen-induced regeneration upon basal and secretory cell proliferation. Prostate 11:339-352.
Guo Y., Sklar G.N., Borkowski A., Kyprianou N. (1997) Loss of the cyclin-dependent kinase inhibitor p27(kip1) protein in human prostate cancer correlates with tumor grade. Clin Cancer Res 3:2269-2274.
Durand B., Gao F.B., Raff M. (1997) Accumulation of the p27/KIP1 cyclin-dependent kinase inhibitor and the timing of oligodendrocyte differentiation. EMBO J 16:306-317.
Hiromura K., Fero M.L., Roberts J.M., Shankland S.J. (1999) Modulation of apoptosis by the cyclin-dependent kinase inhibitor p27. J Clin Invest 103:597-604.
Ophascharoensuk V., Fero M.L., Hughes J., Roberts J.M., Shankland S.J. (1998) The cyclin dependent kinase inhibitor p27 safeguards against inflammatory injury. Nat Med 4:575-580.
Eymin B., Haugg M., Droin N., Sordet O., Dimanche-Boitrel M.-T., Solary E. (1999) p27 induces drug resistance by preventing apoptosis upstream of cytochrome c release and procaspase-3 activation in leukemic cells. Oncogene 18:1411-1418.
Minami S., Ohtani-Fujita N., Igata E., Tamaki T., Sakai T. (1997) Molecular cloning and characterization of the human p27 gene promoter. FEBS Lett 411:1-6.
Sutterluty H., Chatelain E., Marti A., Wirbelauer C., Senften M., Muller U., Krek W. (1999) skp2 promotes p27 degradation and induces S phase in quiescent cells. Nat Cell Biol 1:207-214.
Wirbelauer C., Sutterluty H., Blondel M., Gstaiger M., Peter M., Reymond F., Krek W. (2000) The F-box protein skp2 is a ubiquitylation target of a Cul1-based core ubiquitin ligase complex: Evidence for a role of cul1 in the suppression of skp2 expression in quiescent fibroblasts. EMBO J 19:5362-5375.
Zhang H., Kobayashi R., Galaktionov K., Beach D. (1995) p19Skp1 and p45Skp2 are essential elements of the cyclin A-CDK2 S phase kinase. Cell 82:915-925.
Lu S., Liu M., Epner D.E., Tsai S.Y., Tsai M.J. (1999) Androgen regulation of the cyclin-dependent kinase inhibitor p21 gene through an androgen response element in the proximal promoter. Mol Endocrinol 13:376-384.
Chiarle R., Budel L.M., Skolnik J., Frizzera G., Chilosi M., Corato A., Pizzolo G., Magidson J., Montagnoli A., Pagano M., Maes B., De Wolf-Peeters C., Inghirami G. (2000) Increased proteasome degradation of cyclin-dependent kinase inhibitor p27 is associated with a decreased overall survival in mantle cell lymphoma. Blood 95:619-626.
Fink L., Seeger W., Ermert L., Hanze J., Stahl U., Grimminger F., Kummer W., Bohle R.M. (1998) Real-time quantitative RT-PCR after laser-assisted cell picking. Nat Med 4:1329-1333.
Thomas G.V., Szigeti K., Murphy M., Draetta G., Pagano M., Loda M. (1998) Down-regulation of p27 is associated with development of colorectal adenocarcinoma metastases. Am J Pathol 153:681-687.
Signoretti S., Waltregny D., Dilks J., Isaac B., Lin D., Garraway L., Yang A., McKeon F., Loda M. (2000) p63 is a prostate basal cell marker and is required for prostate development. Am J Pathol 157:1769-1775.
