Skip to main content
SpringerLink
Log in

Effects of selected chemotherapeutic agents on PCNA expression in prostate carcinoma cell lines

  • Original Paper
  • Published:
Urological Research Aims and scope Submit manuscript

Abstract

Bivariate flow cytometric analysis of proliferating cell nuclear antigen (PCNA) was performed on prostate carcinoma cell lines (PC-3, DU-145). For both cell lines 100% methanol fixation provided optimal fluorescence intensity of PCNA. The ratio of PCNA/DNA increased in late G1 through early S/phase, followed by a decrease in mid- and late S and enhancement in G2/M phase. PCNA expression was increased in G2/M phase cells treated for 48 h with vinblastine. A slight decrease in PCNA expression was observed with cyclohexamide treatment. Hydroxyurea induced an increase in S-phase fraction along with enhanced PCNA expression. Methotrexate and Adriamycin had little effect on the cell cycle compartments of PC-3 or DU-145; however, methotrexate decreased PCNA expression, while Adriamycin enhanced it. Cisplatin increased S-phase in both cell lines, increasing PCNA expression in PC-3 and decreasing it in DU-145 cells. The data on the effects of drug treatment point to a dissociation between PCNA expression and S-phase fraction as calculated from the DNA distribution. In some cases, e.g., the cisplatin studies, different effects were obtained in the two different cell lines treated with the same drugs. Whether changes in PCNA expression will provide more useful information than S-phase fraction for evaluation of potential antitumor drugs is not known.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bauer KD (1990) Analysis of proliferation-associated antigens. In: Darzynkiewicz Z, Crissman HA (eds) Flow cytometry. Academic Press, San Diego, Calif, p 235

    Google Scholar 

  2. Bhuyan BK, Groppi VE (1989) Cell-cycle-specific inhibitors. Pharmacol Ther 42:307

    Google Scholar 

  3. Bhyuan BK, Scheidt LG, Day KJ (1972) Cell cycle phase specificity of antitumor agents. Cancer Res 32:398

    Google Scholar 

  4. Bodley A, Liu LF, Israel M, Seshadri R, Koseki Y, Giulani FC, Kirschenbaum S, Silber R, Potmesil M (1989) DNA topoisomerase II-mediated interaction of doxorubin and daunorubicin congeners with DNA. Cancer Res 49:5969

    Google Scholar 

  5. Bolton WE, Mikulka WR, Healy CG, Schmittling RJ, Kenyon NS (1992) Expression of proliferation associated antigens in the cell cycle of synchronized mammalian cells. Cytometry 13:117

    Google Scholar 

  6. Bravo R, Celis JE (1985) Changes in the nuclear distribution of cyclin (PCNA) during S-phase are not triggered by post transitional modifications that are expected to moderately affect its charge. FEBS Lett 185:435

    Google Scholar 

  7. Bravo R, Macdonald-Bravo H (1984) Induction of the nuclear protein “cyclin” in quiescent mouse 3T3 cells stimulated by serum and growth factors: correlation with DAN synthesis. EMBO J 3:177

    Google Scholar 

  8. Bravo R, MacDonald-Bravo H (1987) Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol 105:1549

    Google Scholar 

  9. Bruno S, Crissman HA, Bauer KD, Darzynkiewicz Z (1991) Changes in cell nuclei during S-phase: progressive chromatin condensation and altered expression of the proliferation-associated nuclear protein Ki-67, cyclin (PCNA), p 105 and p34. Exp Cell Res 196:99

    Google Scholar 

  10. Bruno S, Darzynkiewicz Z (1992) Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60. Cell Prolif 25:31

    Google Scholar 

  11. Busch H, Smetana K (1970) The nucleolus. Academic Press, New York

    Google Scholar 

  12. Darzynkiewicz Z, Traganos F, Xue S, Staiano-Coico L, Melamed MR (1981) Rapid analysis of drug effects on the cell cycle. Cytometry 1:279

    Google Scholar 

  13. DelBino G, Darzynkiewicz Z (1991) Camptothecin, teniposide, or 4′-(9-acridinylamino)-3-methanesulfon-m-anisidide, but not mitoxantrone or doxorubin, induces degradation of nuclear DNA in the S-phase of HL-60 cells. Cancer Res 51:1165

    Google Scholar 

  14. Einhorn LH (1983) An overview of chemotherapeutic trials in advanced cancer of the prostate. In: Skinner DG (ed) Urological cancer. Grunne & Stratton, New York, p 89

    Google Scholar 

  15. Elder JS, Gibbons RP (1985) Results of trials of the USA national prostate cancer project. In: Schroeder F, Richards B, (eds) Therapeutic principles in metastatic prostate cancer. Liss, New York, p 221

    Google Scholar 

  16. Elias JM, Qiao L, Heimann A, Engellenner WE, Abel W (1992) Paraffin embedded breast carcinomas for the immunohistochemical study of prognostic factors. J Histotechnol 15:315

    Google Scholar 

  17. Fairman MP (1988) DNA polymerase delta/PCNA: actions and interactions. J Cell Sci 94:117

    Google Scholar 

  18. Garcia RL, Coltrera MD, Gown A (1989) Analysis of proliferative grade using anti-PCNA/cyclin monoclonal antibodies in fixed embedded tissues. Am J Pathol 134:733

    Google Scholar 

  19. Gibbons RP (1987) Prostate Cancer: chemotherapy. Cancer 60:586

    Google Scholar 

  20. Giordano M, Danova M, Pellicciari C, Wilson GD, Mazzini G, Conti AMF, Franchini G, Riccardi A, Romanini MGM (1991) Proliferating cell nuclear antigen (PCNA/cyclin) expression during the cell cycle in normal and leukemic cells. Leuk Res 11:965

    Google Scholar 

  21. Gorczyca W, Bruno S, Melamed MR, Darzynkiewicz Z (1992) Cell cycle-related expression of p 120 nuclear antigen in normal human lymphocytes and in cells of HL-60 and MOLT-4 leukemic lines: effects of methotrexate, camptothecin, and teniposide. Cancer Res 52:3491

    Google Scholar 

  22. Hall PA, Levison DA (1990) Review: assessment of cell proliferation in histological material. J Clin Pathol 43:184

    Google Scholar 

  23. Harper ME, Glynne-Jones E, Goddard L Wilson DW, Matenhelia SS; Conn IG, Peeling WB, Griffiths K (1992) Relationship of proliferating cell nuclear antigen (PCNA) in prostate carcinomas to various clinical parameters. Prostate 20:243

    Google Scholar 

  24. Issacs JT (1985) New principles in the management of metastatic prostate cancer. In: Schroeder F, Richards B (eds) Therapeutic principles in metastatic prostate cancer. Liss, New York, p 383

    Google Scholar 

  25. Kaighn ME, Narayan KS, Ohnuki Y, Lechner JF, Jones LW (1979) Establishment and characterization of a human prostate carcinoma cell line (PC-3). Invest Urol 17:16

    Google Scholar 

  26. Kim SH, Kim JH (1972) Lethal effect of Adriamycin on the division cycle of HeLa cells. Cancer Res 32:323

    Google Scholar 

  27. Landberg G, Roos G (1992) Flow cytometric analysis of proliferation associated nuclear antigens using washless staining of unfixed cells. Cytometry 13:230

    Google Scholar 

  28. Littleton RJ, Baker GM, Soomro IN, Adams RL, Whimster WF (1991) Kinetic aspects of Ki-67 antigen expression in a normal cell line. Virch Arch [Cell Pathol] 60:15

    Google Scholar 

  29. Nemoto R, Uchida K, Shimazui T, Hattori K, Koiso H, Harada M (1989) Immunohistochemical demonstration of S-phase cells by anti-Bromodeoxyuridine monoclonal antibody in human prostate adenocarcinoma. J Urol 141:337

    Google Scholar 

  30. Ohyama S, Yonemura Y, Tsugawa K, Miyazaki I, Tanaka M, Sasaki T (1991) Discrimination of mitotic cells using anti-p 105 monoclonal antibody to analyze the mode of action of etoposide and podophyllotoxin in human gastric cancer cells. Jpn J Cancer Res 82:1258

    Google Scholar 

  31. Qiao L, Pizzolo JG, Gorczyca W, Melamed MR (1993) p 145 expression and effects of differentiation during the cell cycle in HL-60 cell line and in normal human lymphocytes: effects of chemotherapeutic agents. Cytometry 14 [Suppl 6]:61

    Google Scholar 

  32. Qiao L, Pizzolo JG, Kusuda L, Miller TB, Melamed MR (1993) Bivariate flow cytometric analysis of nuclear antigens (PCNA, Ki-67, and p105) in fresh tissues using washless mehtod: preliminary study. Am J Clin Pathol 99:348

    Google Scholar 

  33. Reiner WG, Scott WW; Eggleston JC, Walsh PC (1979) Longterm survival after hormonal therapy for stage D prostate cancer. J Urol 122:183

    Google Scholar 

  34. Rizzo MG, Ottavio L, Travali S, Chang C, Kaminska B, Baserga R (1990) The promoter of the human proliferating cell nuclear antigen (PCNA) is bidirectional. Exp Cell Res 188:286

    Google Scholar 

  35. Roberts JJ, Thomson AJ (1979) The mechanism of action of antitumor platinum compounds. Prog Nucl Acid Res Mol Biol 22:71

    Google Scholar 

  36. Schmenti KJ, Jacobberger JW (1992) Fixation of mammalian cells for flow cytometric evaluation of DNA content and nuclear immunofluorescence. Cytometry 13:48

    Google Scholar 

  37. Silverberg E, Lubera JA (1989) Cancer statistics. CA 39:3

    Google Scholar 

  38. Sinclair WK (1967) Hydroxyurea: effects on Chinese hamster cells grown in culture. Cancer Res 27:297

    Google Scholar 

  39. Stone KR, Michey DD; Wunderli H, Mickey GH, Paulson DF (1978) Isolation of human prostate carcinoma cell line (DU-145). Int J Cancer 21:274

    Google Scholar 

  40. Tobey RA (1972) Effects of cytosine arabinoside, daunomycin, mithramycin, azacytidine, adriamycin, and camptothecin on mammalian cell cycle traverse. Cancer Res 32:2720

    Google Scholar 

  41. Toschi L, Bravo R (1988) Changes in cyclin/PCNA distribution during DNA repair synthesis. J Cell Biol 107:1623

    Google Scholar 

  42. Traganos F (1983) Dihydroantraquinone and related bis (substituted) aminoantraquinones: a novel class of antitumor agents. Pharmacol Ther 22:199

    Google Scholar 

  43. Tranganos F, Kimmel M, Bueti C, Darzynkiewicz Z (1987) Effects of inhibition of RNA or protein synthesis on CHO cell cycle progression. J Cell Physiol 133:277

    Google Scholar 

  44. Yarbro JW (1992) Mechanism of action of hydroxyurea. Semin Oncol 19 [Suppl 9]:1

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, New York Medical College, 10595, Valhalla, NY, USA

    L. Qiao, G. Pizzolo & M. R. Melamed

Authors
  1. L. Qiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Pizzolo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. R. Melamed
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by NIH grants Ro-1-CA 14134

Rights and permissions

Reprints and permissions

About this article

Cite this article

Qiao, L., Pizzolo, G. & Melamed, M.R. Effects of selected chemotherapeutic agents on PCNA expression in prostate carcinoma cell lines. Urol. Res. 22, 171–176 (1994). https://doi.org/10.1007/BF00571846

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00571846

Key words

Search

Navigation