Objective: We describe a new human bladder carcinoma cell line (DAG–1) established from a resected bladder cancer fragment and maintained in culture for more than 5 years and over 300 passages. Methods and Results: Immunological, biochemical and molecular analysis showed that the DAG–1 cells (62 chromosomes) express the cytokeratines 8, 13, 18 and 20 that confirm their epithelial origin as well as numerous cytokine and cytokine receptor mRNAs. They secrete tissue–type plasminogen activator (t–PA), urokinase–type plasminogen activator (u–PA), plasminogen activator inhibitors (PAI–1 and PAI–2), and express u–PA receptors (u–PAR/CD87) at their surface. DAG–1 cells are resistant to TNFα– and IFNγ–induced apoptosis, two cytokines secreted in the urine of Calmette–Guérin bacillus–treated patients and involved in the tumor regression. Conclusion: The DAG–1 cell line is a useful tool, both in vitro and in vivo, to study the progression of bladder tumors and their mechanisms of resistance to immunotherapy in relation with PAI–2 and antioxidant enzymes.

Keywords:
Bladder cell line, Fibrinolytic activity, Resistance to BCG Therapy
1.
Davies B, Waxman J, Wasan H, Abel P, Williams G, Krausz T, Neal D, Thomas D, Handy A, Balkwill F: Levels of matrix metalloproteases in bladder cancer correlate with tumor grade and invasion. Cancer Res 1993;53: 5365–5368.
2.
Gohji K, Fujimoto N, Komiyama T, Fujii A, Ohkawa J, Kamidomo S, Nakajima M: Elevation of serum levels of matrix metalloproteinase–2 and –3 as new predictors of recurrence in patients with urothelial carcinoma. Cancer 1996;78:2379–2387.
3.
Smith K, Fennely JA, Neal DE, Hall RR, Harris AL: Characterization and quantitation of the epidermal growth factor in invasive and superficial bladder tumors. Cancer Res 1989;49: 5810–5814.
4.
Mizutani Y, Okada Y, Terachi T, Kakehi Y, Yoshida O: Serum granulocyte–colony stimulating factor levels in patients with urinary bladder tumor and various urological malignancies. Br J Urol 1995;76:580–585.
5.
William RB: The fibrinolytic system in neoplasia. Semin Thromb Hemost 1996;22:459– 478.
6.
Champelovier P, Valiron O, Jacrot M, Leroux D, Seigneurin D: Selection and characterization of an erythroeosinophilic subclone (LAA–87) and an eosinophilic subclone (LAM–88) from the multipotential cell line LAMA–84. Exp Haematol 1994;18:903–918.
7.
Mitelman F (ed): An International System for Human Cytogenetic Nomenclature (1991) ISCN (1991). Guidelines for Cancer Cytogenetics: Supplement. Basel, Karger, 1995.
8.
Fixe P, Lorgeot V, Le Meur Y, Coupey L, Heymann D, Golard A, Preloran V: Development of enzymoimmunoassays (EIA) for macrophage colony–stimulating factor (M–CSF) and leukemia inhibitory factor (LIF) by using the same capture and signal generating polyclonal antibody. Cytokine 1996;8:586–590.
9.
Granelli–Piperno A, Reich E: A study of proteases and protease–inhibitor complexes in biological fluids. J Exp Med 1978;148:223–234.
10.
Fixe P, Rougier F, Ostyn E, Gachard N, Faucher JL, Praloran V, Denizot Y: Spontaneous and inducible production or macrophage colony–stimulating factor by human bone marrow stromal cells. Eur Cytokine Netw 1997;8: 9–15.
11.
Champelovier P, Richard MJ, Seigneurin D: Autocrine regulation of TPA–induced apoptosis in monoblastic cell line U–937: Role for TNFα, MnSOD and IL–6. Anticancer Res 2000;20:451–458.
12.
Douglas AM, Goss GA, Sutherland RL, Hilton DJ, Berndt MC, Nicola NA, Begley CG: Expression and function of members of the cytokine receptor superfamily on breast cancer cells. Oncogene 1997;14:661–669.
13.
Tachibana M, Miyakawa A, Uchida A, Murai M, Eguchi K, Nakamura K, Kubo A, Hata JI: Granulocyte colony–stimulating factor receptor expression on human transitional cell carcinoma of the bladder. Br J Cancer 1997;75: 1489–1496.
14.
Oka M, Hirose K, Iizuka N, Aoyagi K, Yamamoto K, Abe T, Hazama S, Suzuki T: Cytokine mRNA expression patterns in human esophageal cancer cell lines. J Interferon Cytokine Res 1995;15:1005–1011.
15.
Avalos BR, Gasson JC, Hedvat C, Quan SG, Baldwin GC, Weisbart RH, Williams RE, Golde DW, Di Persio JF: Human granulocyte colony–stimulating factor: Biologic activities and receptor characterization on hematopoietic cells and small cell lung cancer cell lines. Blood 1990;75:851–857.
16.
Savarese DM, Valinski H, Queseberry P, Savarese T: Expression and function of colony–stimulating factors and their receptors in human prostate carcinoma cell lines. Prostate 1998;34:80–85.
17.
Berdel WE, Danhauser–Rield S, Steinhauser G, Winton EF: Various human hematopoietic growth factors (interleukin3, GM–CSF, G–CSF) stimulate clonal growth of nonhematopoietic tumor cells. Blood 1989;73:80–83.
18.
Maher MG, Sapi E, Turner B, Gumbs A, Perrotta PL, Carter D, Kacinski BM, Andaffy BG: Prognostic significance of colony–stimulating factor receptor expression in ipsilateral breast cancer recurrence. Clin Cancer Res 1998;4: 1851–1856.
19.
Tachibana M, Miyakawa A, Tazaki H, Nakamura K, Kubo A, Hata JI, Nishi T, Amano Y: Autocrine growth of transitional cell carcinoma of the bladder induced by granulocyte–colony stimulating factor. Cancer Res 1995; 55:3438–3443.
20.
Filderman AE, Bruckner A, Kacinski BM, Deng N, Remold HG: Macrophage colony–stimulating factor (CSF–1) enhances invasiveness in CSF–1 receptor–positive carcinoma cell lines. Cancer Res 1993;52:3661–3666.
21.
Imao T, Koshida K, Endo Y, Uchibayashi T, Sasaki T, Namiki M: Dominant role of E–cadherin in the progression of bladder cancer. J Urol 1999;161:692–698.
22.
Hudson MA, McReynolds LM: Urokinase and the urokinase receptor: Association with in vitro invasiveness of human bladder cancer cell lines. J Natl Cancer Inst 1997;89:709–717.
23.
George F, Pourreau–Schneider N, Arnoux D, Boutière B, Dussault N, Roux–Dosseto M, Alessi MC, Martin PM, Sampol J: Modulation of tPA, PAI1 and PAI2 antigen and mRNA levels by EGF in the A431 cell line. Blood Coagul Fibrinolysis 1990;1:689–693.
24.
Hasui Y, Marutsuka K, Osada Y, Osada Y: Prognosis value of urokinase–type plasminogen activator in patients with superficial bladder cancer. Urology 1996;47:34–37.
25.
Andreasen PA, Kjoller L, Christensen L, Duffy MJ: The urokinase–type plasminogen activator system in cancer metastasis: A review. Int J Cancer 1997;72:1–22.
26.
Taniguchi K, Koga S, Nishikido M, Yamshita S, Sakuragi T, Kanetake H, Saito Y: Systemic immune response after intravesical instillation of bacille Calmette–Guérin (BCG) for superficial bladder cancer. Clin Exp Immunol 1999; 115:131–135.
27.
Dickinson JL, Bates EJ, Ferrante A, Antalis T: Plasminogen activator inhibitor type 2 inhibits tumor necrosis factor alpha–induced apoptosis. Evidence of an alternative biological function. J Biol Cell 1995;46:27894–27904.
2001
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.