DOCSLIB.ORG

Osteoblast-Derived Factors Induce Androgen-Independent Proliferation and Expression of Prostate-Specific Antigen in Human Prostate Cancer Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Osteoblast-Derived Factors Induce Androgen-Independent Proliferation and Expression of Prostate-Specific Antigen in Human Prostate Cancer Cells 1860 Vol. 10, 1860–1869, March 1, 2004 Clinical Cancer Research Osteoblast-Derived Factors Induce Androgen-Independent Proliferation and Expression of Prostate-Specific Antigen in Human Prostate Cancer Cells Natalie Blaszczyk,1 Bassam A. Masri,4 cotreated with both OCM and androgen. OCM targeted the 1 1 Nasrin R. Mawji, Takeshi Ueda, NH2-terminal domain of the AR. The effect of OCM on transcriptional activity of the AR was inhibited by an anti- Gavan McAlinden,4 Clive P. Duncan,4 1 2 androgen. Neutralizing antibodies to IL-6 blocked prolifer- Nicholas Bruchovsky, Hans-Udo Schweikert, ation and expression of PSA by OCM. 2 3 Doris Schnabel, Edward C. Jones, and Conclusion: Osteoblasts secrete factors, such as IL-6, Marianne D. Sadar1 that cause androgen-independent induction of PSA gene 1Department of Cancer Endocrinology, British Columbia Cancer expression and proliferation of prostate cancer cells by a Agency, Vancouver, British Columbia, Canada; 2Department of mechanism that partially relies on the AR. Identifying such Internal Medicine, University of Bonn, Bonn, Germany; and molecular mechanisms may lead to improved clinical man- 3 4 Anatomical Pathology and Orthopaedic Surgery, Vancouver agement of metastatic prostate cancer. General Hospital, British Columbia, Canada INTRODUCTION ABSTRACT Localized carcinoma of the prostate (CaP) can potentially Purpose: Prostate cancer metastasizes to the skeleton to be cured by surgery or radiation therapy. However, the only form osteoblastic lesions. Androgen ablation is the current treatment available for advanced metastatic disease is the with- treatment for metastatic prostate cancer. This therapy drawal of androgens, which are essential for the survival of is palliative, and the disease will return in an androgen- prostate epithelial cells (for review, see Ref. 1). Clinical re- independent form that is preceded by a rising titer of pros- sponse to androgen ablation therapy is temporary with the tate-specific antigen (PSA). Here, we investigated the possi- ultimate development of androgen-independent disease that is bility that human osteoblasts might secrete factors that preceded by an increasing titer of serum prostate-specific anti- contribute to the emergence of androgen-independent pros- gen (PSA). Unlike most malignancies, up to 85% of CaP me- tate cancer. tastases occur in the bone as osteoblastic (bone forming) lesions. Experimental Design: Primary cultures of human osteo- Osteoblastic lesions are the major cause of CaP-related morbid- blasts were used as a source of conditioned medium (OCM). ity, and mortality and even small, bone-limited tumor burden in Proliferation, expression of androgen-regulated genes, and patients are strongly correlated to cachexia and death (for re- transactivation of the androgen receptor (AR) were moni- view, see Ref. 2). tored in LNCaP human prostate cancer cells in response to Bone is mostly composed of an acellular collagen matrix and OCM using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl- abundant in immobilized growth factors. Dispersed throughout this tetrazolium bromide (MTT) assay, Northern blot analysis, matrix are osteoblasts and osteoclasts, which are the cells respon- and reporter gene constructs. Levels of interleukin-6 (IL-6) sible for bone maintenance. In bone, remodeling osteoclasts de- present in OCM were measured, and its contribution to grade the matrix to release growth factors which stimulate osteo- proliferation and expression of PSA were investigated by blasts to lay down new bone (for review, see Refs. 3 and 4). Many neutralization studies with anti IL-6 antibodies. of these growth factors increase proliferation of prostate cancer Results: OCM increased the proliferation and expres- cells. In vitro studies have shown increased proliferation of prostate sion of PSA at both the protein and RNA levels in LNCaP cancer cells stimulated by conditioned media from or cocultured cells. Synergistic increases in the activities of PSA (6.1 kb)- with osteoblasts, growth factor extracts, and individual growth and pARR3-tk-luciferase reporters were measured in cells factors derived from bone, such as interleukin-6 (IL-6), insulin-like growth factor I (IGF-I) and IGF-II, epidermal growth factor, kera- tinocyte growth factor, bone morphogenic proteins, fibroblast growth factors, transforming growth factor-␤, cytokines, platelet- derived growth factor, vascular endothelial growth factor, and Received 6/17/03; revised 11/19/03; accepted 12/1/03. endothelin-1 (5–13). Many of these growth factors may be in- Grant support: George M. O’Brien Research Center Grant P50 volved in circumventing the need for androgen in advanced disease DK47656 from the NIH (to M. D. S.). The costs of publication of this article were defrayed in part by the by a mechanism involving ligand-independent activation of the payment of page charges. This article must therefore be hereby marked androgen receptor (AR; Refs. 6 and 14–17). advertisement in accordance with 18 U.S.C. Section 1734 solely to The AR is a transcription factor that binds androgens and indicate this fact. regulates gene expression required for normal male sexual de- Requests for reprints: Marianne D. Sadar, Department of Cancer Endocrinology, British Columbia Cancer Agency, 600 West 10th Ave- velopment and maintenance of secondary sex characteristics nue, Vancouver, B. C., V5Z 4E6 Canada. Phone: (604) 877-6036; Fax: (for review, see Ref. 1). It is expressed in the majority of (604) 877-6011; E-mail: [email protected]. prostate cancer tissue specimens, including androgen independ- Downloaded from clincancerres.aacrjournals.org on October 2, 2021. © 2004 American Association for Cancer Research. Clinical Cancer Research 1861 ent or hormone refractory disease, and is therefore a strong All experiments were performed using serum- and phenol red- candidate for mediating androgen resistance (18–20). The AR is free conditions. composed of three domains. Centrally located is the DNA- Culturing of Osteoblast-Like Cells and Preparing binding domain (DBD) that binds to androgen response ele- Osteoblast-Conditioned Medium (OCM)-MEM. Human ments (AREs) in upstream regulatory regions of androgen- osteoblast-like cells were cultured from femoral head trabecular regulated genes, such as PSA. The most COOH-terminal region explants obtained from osteoarthritis patients undergoing hip/ comprises the ligand-binding domain that binds androgens and knee replacement surgery. To minimize possible patient varia- antiandrogens, such as bicalutamide. The ligand-binding do- tion, donors were restricted to males Ͻ 65 years of age. Tra- main contains a weak activation function-2 region and is sepa- becular bone was scraped into bone chips and further processed rated from the DBD by a hinge region, which mediates nuclear with a mortar and pestle. Bone chips were cultured in MEM containing 20% FBS at 37°C in the presence of 5% CO (35). localization. The NH2-terminal domain (NTD) is the most vari- 2 able in sequence homology between species and contains the The outgrowth of the osteoblast-like cells was monitored visu- activation function-1 region required for transactivation (for ally under the microscope with the aid of Gram Safaran staining. review, see Ref. 1). The AR can be activated in an androgen- Cells were characterized using the reverse transcriptase-PCR for independent manner by a number of factors, including IL-6, IGF expression of the following osteoblast markers: (a) type I pro- I, IGF II, keratinocyte growth factor, epidermal growth factor, collagen; (b) alkaline phosphatase; and (c) osteocalcin (35). forskolin, and cAMP, and some factors such as IL-6 and cAMP When the osteoblast-like cells were confluent, they were washed ϫ have been shown to mediate activation via the AR NTD (6, with PBS (3 20 ml) and cultured in 20 ml of serum-free MEM. After 48 h, the OCM was collected and centrifuged or 14–16). Because the bone environment is rich in many of these filtered through 0.22-␮m Nalgene units to remove the cellular growth factors, it has been suggested that bone-derived factors debris before storing at Ϫ80°C until use. OCMs from primary may facilitate survival and progression of CaP to androgen cultures of osteoblasts prepared from different patients were independence by cross-talk with the AR and alternative signal never pooled but rather used individually. Primary cultures of transduction pathways (5, 8, 21). osteoblasts were grown and maintained in Falcon Primaria T75 Clinical and experimental evidence suggests that one of the flasks, with a surface of 75 cm2. The average protein concen- more important factors in prostate cancer progression to andro- tration in OCM was 0.28 Ϯ 0.05 mg/ml (SE). Fibroblast- gen independence and development of osseous metastases is conditioned media were collected and used as reported pre- IL-6. Elevated levels of IL-6 have been found in sera from viously (5). patients who have metastatic and/or hormone refractory disease 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (22–26). IL-6 was first characterized as a modulator of an Bromide (MTT) Assay. LNCaP cells (1 ϫ 104) were plated immune response that is produced by T cells and involved in in 96-well Falcon tissue culture plates in RPMI containing 0.5% hematopoiesis and inflammation (for review, see Ref. 27). How- FBS in a final volume of 0.1 ml. When the cells reached 60% ever, IL-6 has been subsequently shown to be produced by many confluence, usually within 24 h, they were treated with R1881 other cell types, including normal and malignant prostate epi- or OCM collected from preparations of osteoblasts from three thelial cells and bone cells (28–34). It is clear that IL-6 has the individual patients. After 5 days of culture, cell proliferation potential to act in both a paracrine and autocrine manner. Im- was assessed by adding 50 ␮l of 3-(4,5-dimethylthiazol-2-yl)- portantly, IL-6 has been shown to enhance proliferation of 2,5-diphenyltetrazolium bromide dye (1 mg/ml) in serum-free prostate cancer cells, including LNCaP (6, 28), and induce media to the cells.
Load more

Recommended publications
  • P53 and PTEN Expression Contribute to the Inhibition of EGFR Downstream Signaling Pathway by Cetuximab
    Cancer Gene Therapy (2009) 16, 498–507 r 2009 Nature Publishing Group All rights reserved 0929-1903/09 $32.00 www.nature.com/cgt ORIGINAL ARTICLE P53 and PTEN expression contribute to the inhibition of EGFR downstream signaling pathway by cetuximab S Bouali1, A-S Chre´tien1, C Ramacci1, M Rouyer1, S Marchal2, T Galenne3, P Juin3, P Becuwe4 and J-L Merlin1 1Centre Alexis Vautrin, Unite´ de Biologie des Tumeurs, EA ‘‘Predicther’’ Nancy Universite´, Vandœuvre-le`s- Nancy cedex, France; 2Centre Alexis Vautrin, CRAN UMR 7039 CNRS-UHP-INPL, Vandœuvre-le`s-Nancy cedex, France; 3INSERM U601, CHU de Nantes, Institut de Biologie, Nantes, France and 4Laboratoire de Biologie Cellulaire, Faculte´ des Sciences, EA ‘‘Predicther’’ Nancy Universite´, Vandœuvre-le`s-Nancy, France Cetuximab (Erbitux) is an anti-epidermal growth factor receptor (EGFR) monoclonal antibody whose activity is related to the inhibition of EGFR downstream signaling pathways. P53 and phosphatase and tensin homologue deleted on chromosome 10 (PTEN) have been reported to control the functionality of PI3K/AKT signaling. In this study we evaluated whether reintroducing P53 using non-viral gene transfer enhances PTEN-mediated inhibition of PI3K/AKT signaling by cetuximab in PC3 prostate adenocarcinoma cell line bearing p53 and pten mutations. Signaling phosphoproteins expression was analyzed using Bio-Plex phosphoprotein array and western blot. Apoptosis induction was evaluated from BAX expression, caspase-3 activation and DNA fragmentation analyses. The results presented show that p53 and pten gene transfer additionally mediated cell growth inhibition and apoptosis induction by restoral of signaling functionality, which enabled the control of PI3K/AKT and MAPKinase signaling pathways by cetuximab in PC3 cells.
    [Show full text]
  • Indirect Antitumor Effects of Bisphosphonates on Prostatic Lncap Cells Co-Cultured with Bone Cells
    ANTICANCER RESEARCH 29: 1089-1094 (2009) Indirect Antitumor Effects of Bisphosphonates on Prostatic LNCaP Cells Co-cultured with Bone Cells TOMOAKI TANAKA, HIDENORI KAWASHIMA, KEIKO OHNISHI, KENTARO MATSUMURA, RIKIO YOSHIMURA, MASAHIDE MATSUYAMA, KATSUYUKI KURATSUKURI and TATSUYA NAKATANI Department of Urology, Osaka City University Graduate School of Medicine, Osaka 545-8585, Japan Abstract. Bisphosphonates are strong inhibitors of effectively inhibit bone absorption are available for osteoclastic bone resorption in both benign and malignant reducing the risk of skeletal complications in bone bone diseases. The nitrogen-containing bisphosphonates metastases of prostate cancer as well as those of other solid (N-BPs) have strong cytotoxicity via inhibition of protein tumors characterized by osteoclastic lesions. prenylation in the mevalonate pathway, and also demonstrate Bisphosphonates are potent inhibitors of osteoclast activity direct cytostatic and proapoptotic effects on prostate cancer and survival, thereby reducing osteoclast-mediated bone cells. We confirmed the usefulness of a co-culture system absorption (1, 4-6). The chemical structure of bisphosphonates comprised of prostatic LNCaP cells, ST2 cells (mouse- contributes to their mechanism of action: Non-nitrogen- derived osteoblasts) and MLC-6 cells (mouse-derived containing compounds (e.g. etidronate, clodronate) are osteoclasts) in vitro. N-BPs (pamidronate and zoledronic metabolized to cytotoxic analogues of ATP (7), while newer acid) inhibited both androgen receptor transactivation and nitrogen-containing bisphosphonates (N-BPs; pamidronate, tumor cell proliferation by suppressing the activities of both ibandronate, zoledronic acid) have strong cytotoxic effects on osteoclasts and osteoblasts with low-dose exposure. This cells via inhibition of protein prenylation in the mevalonate indirect inhibition of prostate cancer cells via bone cells pathway (8, 9).
    [Show full text]
  • Prostate Cancer Stromal Cells and Lncap Cells Coordinately Activate
    Endocrine-Related Cancer (2009) 16 1139–1155 Prostate cancer stromal cells and LNCaP cells coordinately activate the androgen receptor through synthesis of testosterone and dihydrotestosterone from dehydroepiandrosterone Atsushi Mizokami, Eitetsu Koh, Kouji Izumi, Kazutaka Narimoto, Masashi Takeda, Seijiro Honma1, Jinlu Dai 2, Evan T Keller 2 and Mikio Namiki Department of Integrative Cancer Therapy and Urology, Kanazawa University Graduate School of Medical Sciences, 13-1 Takaramachi, Kanazawa City 920-8640, Japan 1Teikoku Hormone MFG. Co., 1604 Shimosakunobe, Takatsu-ku, Kawasaki, Kanagawa 213, Japan 2Departments of Urology and Pathology, University of Michigan, Ann Arbor, Michigan, USA (Correspondence should be addressed to A Mizokami; Email: [email protected]) Abstract One of the mechanisms through which advanced prostate cancer (PCa) usually relapses after androgen deprivation therapy (ADT) is the adaptation to residual androgens in PCa tissue. It has been observed that androgen biosynthesis in PCa tissue plays an important role in this adaptation. In the present study, we investigated how stromal cells affect adrenal androgen dehydroepian- drosterone (DHEA) metabolism in androgen-sensitive PCa LNCaP cells. DHEA alone had little effect on prostate-specific antigen (PSA) promoter activity and the proliferation of LNCaP cells. However, the addition of prostate stromal cells or PCa-derived stromal cells (PCaSC) increased DHEA-induced PSA promoter activity via androgen receptor activation in the LNCaP cells. Moreover, PCaSC stimulated the proliferation of LNCaP cells under physiological concentrations of DHEA. Biosynthesis of testosterone or dihydrotestosterone from DHEA in stromal cells and LNCaP cells was involved in this stimulation of LNCaP cell proliferation. Androgen biosynthesis from DHEA depended upon the activity of various steroidogenic enzymes present in stromal cells.
    [Show full text]
  • Cell Line Lncap'
    ICANCER RESEARCH58. 76-83. January 1. 1998J Caspase-7 Is Activated during Lovastatin-induced Apoptosis of the Prostate Cancer Cell Line LNCaP' Marco Marcelli,2 Glenn R. Cunningham, S. Joe Haidacher, Sebastian J. Padayatty, Lydia Sturgis, Carolina Kagan, and Larry Denner Departments of Medicine (M. M.. G. R. C., S. J. H.. S. f. P.1 and Cell Biology [M. M., G. R. C.], Veterans Affairs Medical Center and Baylor College of Medicine, Department of Cell Biology (L S., C K., L DI, Texas Biotechnology C'orporation, Houston, Texas 77030 ABSTRACT androgen-dependent and androgen-independent cells even before therapy (4). The initial response following androgen ablation is The goals of this work were to establish a reproducible and effective thought to be due to the induction of apoptosis of androgen-dependent model of apoptosis in a cell line derived from advanced prostate cancer prostate cancer cells (5). Because androgen-independent cells are and to study the role of the caspase family of proteases in mediating insensitive to androgen ablation, they remain alive even in the absence apoptosis in this system. The study involved the use of the prostate cancer cell line LNCaP. Apoptosis was induced using the hydroxymethyl glutaryl of androgens (6). Hence, after the initial response to androgen ablation CoA reductase inhibitor, lovastatin, and was evaluated by agarose gel due to the death of androgen-dependent cells, the tumor progresses to electrophoresis of genomic DNA, morphological criteria, and terminal an androgen-independent state, and the patient is no longer curable deoxynucleotidyl traasferase-mediated nick end labeling. Caspases were (7).
    [Show full text]
  • On the Biological Properties of Prostate Cancer Cells by Modulation of Inflammatory and Steroidogenesis Pathway Genes
    International Journal of Molecular Sciences Article The Impact of Ang-(1-9) and Ang-(3-7) on the Biological Properties of Prostate Cancer Cells by Modulation of Inflammatory and Steroidogenesis Pathway Genes Kamila Domi ´nska 1,* , Karolina Kowalska 2 , Kinga Anna Urbanek 2 , Dominika Ewa Habrowska-Górczy ´nska 2 , Tomasz Och˛edalski 1 and Agnieszka Wanda Piastowska Ciesielska 2 1 Department of Comparative Endocrinology, Medical University of Lodz, Zeligowskiego 7/9, 90-752 Lodz, Poland; [email protected] 2 Department of Cell Cultures and Genomic Analysis, Medical University of Lodz, Zeligowskiego 7/9, 90-752 Lodz, Poland; [email protected] (K.K.); [email protected] (K.A.U.); [email protected] (D.E.H.-G.); [email protected] (A.W.P.C.) * Correspondence: [email protected] Received: 12 July 2020; Accepted: 26 August 2020; Published: 28 August 2020 Abstract: The local renin–angiotensin system (RAS) plays an important role in the pathophysiology of the prostate, including cancer development and progression. The Ang-(1-9) and Ang-(3-7) are the less known active peptides of RAS. This study examines the influence of these two peptide hormones on the metabolic activity, proliferation and migration of prostate cancer cells. Significant changes in MTT dye reduction were observed depending on the type of angiotensin and its concentration as well as time of incubation. Ang-(1-9) did not regulate the 2D cell division of either prostate cancer lines however, it reduced the size of LNCaP colonies formed in soft agar, maybe through down-regulation of the HIF1a gene.
    [Show full text]
  • Active Estrogen Synthesis and Its Function in Prostate Cancer-Derived Stromal Cells
    ANTICANCER RESEARCH 35: 221-228 (2015) Active Estrogen Synthesis and its Function in Prostate Cancer-derived Stromal Cells KAZUAKI MACHIOKA1, ATSUSHI MIZOKAMI1, YURI YAMAGUCHI2, KOUJI IZUMI1, SHINICHI HAYASHI3 and MIKIO NAMIKI1 1Department of Integrative Cancer Therapy and Urology, Kanazawa University Graduate School of Medical Sciences, Ishikawa, Japan; 2Division of Breast Surgery, Saitama Cancer Center, Saitama, Japan; 3Department of Molecular and Functional Dynamics, Graduate School of Medicine, Tohoku University, Sendai, Japan Abstract. Background: It remains unclear whether estrogen Castration-naïve prostate cancer (PCa) develops into is produced in prostate cancer (PCa) and how it functions in castration-resistant prostate cancer (CRPC) during androgen PCa. Materials and Methods: To examine the production of deprivation therapy (ADT) by various mechanisms. estrogen in PCa cells, the concentration of estrogen in the Especially, the androgen receptor (AR) signaling axis plays a medium in which LNCaP cells and PCa-derived stromal cells key role in this specific development. PCa cells mainly adapt (PCaSC) were co-cultured, was measured by liquid themselves to the environment of lower androgen chromatography-mass spectrometry/mass spectrometry (LC- concentrations and change into androgen-hypersensitive cells MS/MS), while aromatase (CYP19) mRNA expression was or androgen-independent cells. Androgens of adrenal origin confirmed by real-time polymerase chain reaction (RT-PCR) and their metabolites synthesized in the microenvironment in methods. To verify whether estrogen is synthesized from an intracrine/paracrine fashion act on surviving PCa cells and testosterone in PCaSC functions, PCaSC were co-cultured secrete prostate specific antigen (PSA). Therefore, new with breast cancer MCF-7-E10 cells, which were stably- recently developed medicines, abiraterone acetate that transfected with ERE-GFP, in the presence of testosterone.
    [Show full text]
  • Cell Adhesion and Chemotaxis in Prostate Cancer Metastasis to Bone: a Minireview
    Prostate Cancer and Prostatic Diseases (2000) 3, 6±12 ß 2000 Macmillan Publishers Ltd All rights reserved 1365±7852/00 $15.00 www.nature.com/pcan Review Cell adhesion and chemotaxis in prostate cancer metastasis to bone: a minireview CR Cooper1* & KJ Pienta1 1University of Michigan Comprehensive Cancer Center, Department of Internal Medicine, Ann Arbor, MI 48109, USA Bone metastasis is a common phenomenon in patients with advanced prostate cancer. The molecular and cellular mechanisms involved in this process are not well understood. Past reviews on this subject primarily focused on prostate tumor growth in the bone marrow and the effects this growth has on bone homeostasis (ie osteoblastic and osteolytic). Cell chemotaxis and adhesion are also important for site-speci®c metastasis. In this review we have focused on chemotactic and cell adhesion molecules potentially involved in prostate cancer metastasis to bone. In addition, recently developed animal models for prostate cancer metastasis to bone are discussed. Prostate Cancer and Prostatic Diseases (2000) 3, 6±12. Keywords: cell adhesion molecules; cytokines; human bone marrow endothelial cells; chemoattractants; prostate cancer cells Introduction cells.7 The inactivation of parathyroid hormone-related protein (PTH-rp) by prostate-speci®c antigen may pro- Prostate cancer bone metastasis is a major clinical prob- mote the osteoblastic reaction seen in prostate cancer lem and is commonly associated with intractable pain, patients. Yoneda has proposed that the minor osteolytic pathological
    [Show full text]
  • A Novel Anti-Androgen, Compound 30, Suppresses Castration-Resistant and MDV3100-Resistant Prostate Cancer Growth in Vitro and in Vivo
    Author Manuscript Published OnlineFirst on March 14, 2013; DOI: 10.1158/1535-7163.MCT-12-0798 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. A novel anti-androgen, Compound 30, suppresses castration-resistant and MDV3100-resistant prostate cancer growth in vitro and in vivo Hidetoshi Kuruma1, Hiroaki Matsumoto1, Masaki Shiota1, Jennifer Bishop, Francois Lamoureux1, Christian Thomas1, David Briere2, Gerrit Los2, Martin Gleave1, Andrea Fanjul2 and Amina Zoubeidi1 1The Vancouver Prostate Centre, 2660 Oak Street, Vancouver, BC, Canada, V6H 3Z6 2Pfizer Oncology Research Unit, Worldwide Research & Development, 10646 Science Center Drive, San Diego, California, United States, 92121. Running Title: Efficacy of Compound 30 in MDV3100-resistant prostate cancer Key words: Castration-resistant prostate cancer, anti-androgen, MDV3100, Compound 30, androgen receptor, MR49F Financial support: Pfizer Global Research & Development (United States) and the Prostate Cancer Foundation USA (A. Zoubeidi) Corresponding author: Amina Zoubeidi, Ph.D. The Vancouver Prostate Centre, Department of Urologic Science University of British Columbia: 2660 Oak Street, Vancouver, BC, V6H 3Z6, Canada. [email protected] Disclosure: A. Fanjul, D. Briere and G. Los are employee of Pfizer. The other authors disclosed no potential conflicts of interest. Manuscript Type: Chemical (Small Molecule) Discovery and Clinical Development Total number of figures: 5 Word Count: 4,937 1 Downloaded from mct.aacrjournals.org on September 29, 2021. © 2013 American Association for Cancer Research. Author Manuscript Published OnlineFirst on March 14, 2013; DOI: 10.1158/1535-7163.MCT-12-0798 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
    [Show full text]
  • Repression of Cell Proliferation and Androgen Receptor Activity in Prostate Cancer Cells by 2’-Hydroxyflavanone
    ANTICANCER RESEARCH 33: 4453-4462 (2013) Repression of Cell Proliferation and Androgen Receptor Activity in Prostate Cancer Cells by 2’-Hydroxyflavanone MITSUO OFUDE1, ATSUSHI MIZOKAMI1, MISAKO KUMAKI1, KOUJI IZUMI1, HIROYUKI KONAKA1, YOSHIFUMI KADONO1, YASUHIDE KITAGAWA1, MINKYOUNG SHIN1, JIAN ZHANG2, EVAN T. KELLER3 and MIKIO NAMIKI1 1Department of Integrative Cancer Therapy and Urology, Kanazawa University Graduate School of Medical Sciences, Kanazawa, Ishikawa, Japan; 2Center for Translational Medical Research, Guangxi Medical University, Guangxi, P.R. China; 3Department of Urology, School of Medicine, University of Michigan, Ann Arbor, MI, U.S.A. Abstract. Background: Prevention of the development of line hormonal therapy using medical castration, such as castration-resistant from hormone-naïve prostate cancer is luteinizing hormone releasing hormone (LH-RH) agonists, an important issue in maintaining the quality of life of the LH-RH antagonist, and antiandrogen for advanced PCa. patients. We explored the effect of 2’-hydroxyflavanone on After an initial response to ADT, however, PCa eventually proliferation and androgen responsiveness using prostate loses responsiveness to ADT and progresses into what is cancer cell lines. Materials and Methods: To investigate the termed castration-resistant PCa (CRPC). effect of 2’-hydroxyflavanone on proliferation, prostate The AR axis also plays an important part in the process cancer cells were treated with 2’-hydroxyflavanone. when hormone- sensitive PCa become CRPC. Although Androgen-responsiveness in LNCaP cells was confirmed by serum testosterone decreases to less than 5% before starting luciferase assay after transfection of luciferase reporter ADT, PCa adapts to low serum testosterone level by several driven by prostate specific antigen promoter. To detect mechanisms.
    [Show full text]
  • Human Prostate Cancer in a Clinically Relevant Xenograft Mouse Model: Identification of B(1,6)-Branched Oligosaccharides As a Marker of Tumor Progression
    Published OnlineFirst January 18, 2012; DOI: 10.1158/1078-0432.CCR-11-2900 Clinical Cancer Imaging, Diagnosis, Prognosis Research Human Prostate Cancer in a Clinically Relevant Xenograft Mouse Model: Identification of b(1,6)-Branched Oligosaccharides as a Marker of Tumor Progression Tobias Lange1, Sebastian Ullrich1, Imke Muller€ 1, Michael F. Nentwich1,2, Katrin Stubke€ 1, Susanne Feldhaus1, Christine Knies1, Olaf J.C. Hellwinkel3, Robert L. Vessella7, Claudia Abramjuk8, Mario Anders4, Jennifer Schroder-Schwarz€ 4, Thorsten Schlomm3,6, Hartwig Huland6, Guido Sauter5, and Udo Schumacher1 Abstract Purpose: To establish xenograft mouse models of metastatic and nonmetastatic human prostate cancer and to apply these models to the search for aberrant glycosylation patterns associated with tumor progression in vivo and in patients. Experimental Design: Prostate cancer cells (LNCaP, PC-3, LuCaP 23.1, and DU-145) were xeno- À À À À grafted subcutaneously into immunodeficient pfp / /rag2 / mice. Tumor growth and metastasis formation were quantified and as altered glycosylation patterns have been associated with metastasis formation in several other malignancies, prostate cancer cells were profiled by a quantitative real-time PCR (qRT-PCR) glycosylation array and compared with normal human prostate cells. The activity of upregulated glycosyltransferases was analyzed by their sugar residues end products using lectin histochemistry on primary tumors and metastases in the animal experiments and on 2,085 clinical samples. Results: PC-3 cells produced the largest number of spontaneous lung metastases, followed by LNCaP and LuCaP 23.1, whereas DU-145 was nonmetastatic. qRT-PCR revealed an upregulation of b1,6-N- acetylglucosaminyltransferase-5b (Mgat5b) in all prostate cancer cell lines.
    [Show full text]
  • Androgen Receptor As a Licensing Factor for DNA Replication in Androgen-Sensitive Prostate Cancer Cells
    Androgen receptor as a licensing factor for DNA replication in androgen-sensitive prostate cancer cells Ivan V. Litvinov*†, Donald J. Vander Griend*‡, Lizamma Antony*, Susan Dalrymple*, Angelo M. De Marzo‡§, Charles G. Drake‡¶, and John T. Isaacs*†‡ʈ *Chemical Therapeutics Program, ¶Division of Immunology and Hematopoiesis, The Sidney Kimmel Comprehensive Cancer Center, †Cellular and Molecular Medicine Graduate Program, §Department of Pathology, and the ‡Brady Urological Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231 Edited by Owen N. Witte, University of California, Los Angeles, CA, and approved August 11, 2006 (received for review April 16, 2006) Androgen receptor (AR) protein expression and function are critical AR-stimulated growth (11). As part of this malignant conversion, for survival and proliferation of androgen-sensitive (AS) prostate AR undergoes a molecular switch from its ability to suppress cancer cells. Besides its ability to function as a transcription factor, proliferation of normal prostatic epithelia to directly stimulating the experimental observations suggest that AR becomes a licensing proliferation of prostate cancer cells (4, 11, 12). Such a molecular factor for DNA replication in AS prostate cancer cells and thus must switch requires gain-of-function changes in AR signaling. Recent be degraded during each cell cycle in these cells to allow reinitia- studies documented that one such gain-of-function event involves tion of DNA replication in the next cell cycle. This possibility was DNA rearrangement such that the androgen response element is tested by using the AS human prostate cancer cell lines, LNCaP, translocated to confer androgen responsiveness upon promoters for CWR22Rv1, and LAPC-4.
    [Show full text]
  • Switch from Antagonist to Agonist of the Androgen Receptor Blocker Bicalutamide Is Associated with Prostate Tumour Progression I
    British Journal of Cancer (1999) 81(2), 242–251 © 1999 Cancer Research Campaign Article no. bjoc.1999.0684 Switch from antagonist to agonist of the androgen receptor blocker bicalutamide is associated with prostate tumour progression in a new model system Z Culig1,*, J Hoffmann4,*, M Erdel2, IE Eder1, A Hobisch1, A Hittmair3, G Bartsch1, G Utermann2, MR Schneider4, K Parczyk4 and H Klocker1 Departments of 1Urology, 2Medical Biology and Human Genetics and 3Pathology, University of Innsbruck, Anichstraße 35, A-6020 Innsbruck, Austria; 4Research Laboratories of Schering AG, D-13342 Berlin, Germany Summary Advanced prostate cancer is treated by androgen ablation and/or androgen receptor (AR) antagonists. In order to investigate the mechanisms relevant to the development of therapy-resistant tumours, we established a new tumour model which closely resembles the situation in patients who receive androgen ablation therapy. Androgen-sensitive LNCaP cells were kept in androgen-depleted medium for 87 passages. The new LNCaP cell subline established in this manner, LNCaP-abl, displayed a hypersensitive biphasic proliferative response to androgen until passage 75. Maximal proliferation of LNCaP-abl cells was achieved at 0.001 nM of the synthetic androgen methyltrienolone (R1881), whereas 0.01 nM of this compound induced the same effect in parental cells. At later passages (> 75), androgen exerted an inhibitory effect on growth of LNCaP-abl cells. The non-steroidal anti-androgen bicalutamide stimulated proliferation of LNCaP-abl cells. AR protein expression in LNCaP-abl cells increased approximately fourfold. The basal AR transcriptional activity was 30-fold higher in LNCaP-abl than in LNCaP cells. R1881 stimulated reporter gene activity in LNCaP-abl cells even at 0.01 nM, whereas 0.1 nM of R1881 was needed for induction of the same level of reporter gene activity in LNCaP cells.
    [Show full text]