Investigational New Drugs

, Volume 19, Issue 4, pp 283–291 | Cite as

Changes in Prostate-Specific Antigen (PSA) Level Correlate with Growth Inhibition of Prostate Cancer Cells Treated in Vitro with a Novel Anticancer Drug, Irofulven

  • Barbara A. Woynarowska
  • Arlene L. Higdon
  • Ruben M. Muñoz
  • Perry Bushong
  • Stephen J. Waters


Irofulven (hydroxymethylacylfulvene,HMAF, MGI 114) is a novel agent withalkylating activity and a potent inducer ofapoptosis. It is currently undergoing PhaseII clinical trials for several tumor types,including hormone-refractory prostatecancer. Reduction of serumprostate-specific antigen (PSA) levels hasbeen proposed as a generally usefulendpoint for evaluating the antitumorefficacy of treatments for prostate cancer.However, the utility of PSA as a marker oftumor cell burden could be compromised, ifdrugs directly affected PSA secretionand/or expression. In these studies, weevaluated the effects of irofulven on PSAprotein and mRNA levels during the courseof treatment of prostate tumor cells in vitro. Therate of PSA secretion(normalized per equal cell number) bycontrol and drug treated cells was similar,as determined by a solid phase, two-siteimmunoradiometric assay. Consistent withthe lack of effect of irofulven on PSAprotein level, the drug does not appear toaffect the expression of PSA mRNA (on a percell basis) as assessed by RT-PCR. Thus,changes in PSA secretion and expressionappear to reflect irofulven-induced cellgrowth inhibition rather than reflecting adirect effect of the drug on PSA. Theseresults suggest that PSA should be areasonable marker of tumor burden inirofulven-treated prostate cancerpatients.

Irofulven HMAF prostate specific antigen 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Kelner MJ, McMorris TC, Taetle R: Preclinical evaluation of illudins as anticancer agents: basis for selective cytotoxicity. J Natl Cancer Inst 82: 1562–1565, 1990Google Scholar
  2. 2.
    MacDonald JR, Muscoplat CC, Dexter DL, Mangold GL, Chen SF, Kelner MJ, McMorris TC: Preclinical antitumor activity of 6-hydroxymethylacylfulvene, a semisynthetic derivative of the mushroom toxin illudin S. Cancer Res 57: 279–283, 1997Google Scholar
  3. 3.
    Mangold G, MacDonald JR, Von Hoff D: Antitumor activity of MGI 114 against two human prostate tumor xenograft models. In: 10th NCI-EORTC Symposium on New Drugs in Cancer Therapy, Amsterdam, Netherlands. 1998, pp. 37.Google Scholar
  4. 4.
    Woynarowski JM, Napier C, Koester SK, Chen SF, Troyer DA, Chapman W, MacDonald JR: Effects on DNA integrity and apoptosis induction by a novel antitumor sesquiterpene drug, 6-hydroxymethylacylfulvene (HMAF, MGI 114). Biochem Pharmacol 54: 1181–1193, 1997Google Scholar
  5. 5.
    Herzig MCS, Arnett B, MacDonald JR, Woynarowski JM: Drug uptake and cellular targets of hydroxymethylacylfulvene (HMAF). Biochem Pharmacol 58: 217–225, 1999Google Scholar
  6. 6.
    Woynarowska B, Woynarowski JM, Higdon AL, Muñoz R: Induction of apoptosis in prostate tumor cells by hydroxymethylacylfulvene (HMAF) and oxaliplatin. Proc Amer Assoc Cancer Res 40: 488, 1999Google Scholar
  7. 7.
    Woynarowska BA, Woynarowski JM, Herzig MCS, Roberts K, Higdon, AL, MacDonald JR: Differential Cytotoxicity and Induction of Apoptosis in Tumor and Normal Cells by Hydroxymethylacylfulvene (HMAF). Biochem Pharmacol 59: 1217–1226, 2000Google Scholar
  8. 8.
    Woynarowski JM, Koester S, Woynarowska B, Arnett B, Trevino AV, Chan D, Higdon A, Muñoz R, Herzig MCS, Faivre S: Is drug-induced DNA damage sufficient for apoptosis induction? Proc Amer Assoc Cancer Res 40: 736, 1999Google Scholar
  9. 9.
    Lilja H: A kallikrein-like serine protease in prostatic fluid cleaves the predominant siminal vesicle protein. J Clin Invest 76: 1899–1903, 1985Google Scholar
  10. 10.
    Stamey TA, Yang N, Hay AR, McNeal JE, Freiha FS, Redwine E: Prostate-specific antigen as a serum marker for adenocarcinoma of the prostate. N Engl J Med 317: 909–916, 1987Google Scholar
  11. 11.
    Gleave ME, Hsieh JT, Wu HC, von Eschenbach AC, Chung LW: Serum prostate specific antigen levels in mice bearing human prostate LNCaP tumors are determined by tumor volume and endocrine and growth factors. Cancer Res 52: 1598–1605, 1992Google Scholar
  12. 12.
    Recker F: Prostate specific antigen in the diagnosis of organ confined curable prostate cancer. Schweiz Med Wochenschr 126: 1881–1890, 1996Google Scholar
  13. 13.
    Cama C, Olsson CA, Raffo AJ, Perlman H, Buttyan RKOT, McMahon D, Benson MC, Katz AE: Molecular staging of prostate cancer. II. A comparison of the application of an enhanced reverse transcriptase polymerase chain reaction assay for prostate specific antigen versus prostate specific membrane antigen, J Urol 153: 1373–1378, 1995Google Scholar
  14. 14.
    Berner A, Waere H, Nesland JM, Paus E, Danielsen HE, Fossa SD: DNA ploidy, serum prostate specific antigen, histological grade and immunohistochemistry as predictive parameters of lymph node metastases in T1-T3/M0 prostatic adenocarcinoma. Br J Urol 75: 26–32, 1995Google Scholar
  15. 15.
    Kardamakis D: Tumour serum markers: Clinical and economical aspects. Anticancer Res 16: 2285–2288, 1996Google Scholar
  16. 16.
    Babaian RJ, Camps JL, Frangos DN, Ramirez EI, Tenney DM, Hassell JS et al: Monoclonal prostate-specific antigen in untreated prostatic cancer. Relationship to clinical stage and grade. Cancer 67: 2200–2206, 1991Google Scholar
  17. 17.
    Oesterling JE: Prostate-specific antigen: A valuable clinical tool. Oncology 5: 107–128, 1991Google Scholar
  18. 18.
    Murphy GP, Barren RJ, Erickson SJ, Bowes VA, Wolfert RL, Bartsch G, Klocker H, Pointner J, Reissigl A, McLeod DG, Douglas T, Morgan T, Kenny GM, Ragde H, Boynton AL, Holmes EH: Evaluation and comparison of two new prostate carcinoma markers – Free-prostate specific antigen and prostate specific membrane antigen. Cancer 78: 809–818, 1996Google Scholar
  19. 19.
    Raviv G, Zlotta AR, Janssen T, Descamps F, Vanegas JP, Verhest A, Schulman CC: Do prostate specific antigen and prostate specific antigen density enhance the detection of prostate carcinoma after initial diagnosis of prostatic intraepithelial neoplasia without concurrent carcinoma? Cancer 77: 2103–2108, 1996Google Scholar
  20. 20.
    Sweat SD, Pacelli A, Bergstralh EJ, Slezak JM, Cheng L, Bostwick DG: Androgen receptor expression in prostate cancer lymph node metastases is predictive of outcome after surgery. J Urol 161: 1233–1237, 1999Google Scholar
  21. 21.
    Cutinha PE, Potts KL, Rosario DJ, Hastie KT, Moore KTH, Chapple CR: A prospective audit of the use of a prostate clinic. Br J Urol 78: 733–736, 1996Google Scholar
  22. 22.
    Sokoloff MH, Tso CL, Kaboo R, Nelson S, Ko J, Dorey F, Figlin RA, Pang S, DeKernion J, Belldegrun A: Quantitative polymerase chain reaction does not improve preoperative prostate cancer staging: A clinicopathological molecular analysis of 121 patients. J Urol 156: 1560–1566, 1996Google Scholar
  23. 23.
    Zagars GK, von Eschenbach AC: Prostate-specific antigen. An important marker for prostate cancer treated with external beam radiation therapy. Cancer 72: 538–548, 1993Google Scholar
  24. 24.
    Critz FA, Levinson AK, Williams WH, Holladay DA: Prostate-specific antigen nadir: The optimum level after irradiation for prostate cancer. J Clin Oncol 14: 2893–2900, 1996Google Scholar
  25. 25.
    Hart KB, Porter AT: A rational approach to the treatment of prostate cancer with radiation therapy: lessons for the future. Seminars in Oncology 24: 745–755, 1997Google Scholar
  26. 26.
    Murphy GP: Review of phase II hormone refractory prostate cancer trials. Urology 54: 19–21, 1999Google Scholar
  27. 27.
    Scher HI, Mazumdar M, Kelly WK: Clinical trials in relapsed prostate cancer: Defining the target. Jnci 88: 1623–1634, 1996Google Scholar
  28. 28.
    Beedassy A, Cardi G: Chemotherapy in advanced prostate cancer. Semin Oncol 26: 428–438, 1999Google Scholar
  29. 29.
    Bolla M, Van Poppel H: Prostate cancer. An insight into screening and local treatment. Eur J Cancer [A] 32A: 1434–1435, 1996Google Scholar
  30. 30.
    Montironi R, Bartels PH, Thompson D, Diamanti L, Prete E: Androgen-deprived prostate adenocarcinoma: Evaluation of treatment-related changes versus no distinctive treatment effect with a Bayesian belief network – A methodological approach. Eur Urol 30: 307–315, 1996Google Scholar
  31. 31.
    Gao CL, Dean RC, Pinto A, Mooneyhan R, Connelly RR, McLeod DG, Srivastava S, Moul JW: Detection of circulating prostate specific antigen expressing prostatic cells in the bone marrow of radical prostatectomy patients by sensitive reverse transcriptase polymerase chain reaction. J Urol 161: 1070–1076, 1999Google Scholar
  32. 32.
    Gomella LG, Raj GV, Moreno JG: Reverse transcriptase polymerase chain reaction for prostate specific antigen in the management of prostate cancer. J Urol 158: 326–337, 1997Google Scholar
  33. 33.
    Hayward SW, Dahiya R, Cunha GR, Bartek J, Deshpande N, Narayan P: Establishment and characterization of an immortalized but non-transformed human prostate epithelial cell line: BPH-1. In Vitro Cell Dev Biol Anim 31: 14–24, 1995Google Scholar
  34. 34.
    Makarovskiy AN, Ackerley W, III, Wojcik L, Halpert GK, Stein BS, Carreiro MP, Hixson DC: Application of immunomagnetic beads in combination with RT-PCR for the detection of circulating prostate cancer cells. J Clin Lab Anal 11: 346–350, 1997Google Scholar
  35. 35.
    Wood DP, Jr, Banks ER, Humphreys S, McRoberts JW, Rangnekar VM: Identification of bone marrow micrometastases in patients with prostate cancer. Cancer 74: 2533–2540, 1994Google Scholar
  36. 36.
    Mannello F, Malatesta M, Gazzanelli G: Detection of circulating prostate specific antigen expressing prostatic cells in the bone marrow of radical prostatectomy patients by sensitive reverse transcriptase polymerase chain reaction. J Urol 163: 253–253, 2000Google Scholar
  37. 37.
    Lange PH, Vessella RL: Detection of circulating prostate specific antigen expressing prostatic cells in the bone marrow of radical prostatectomy patients by sensitive reverse transcriptase polymerase chain reaction – Comment. J Urol 161: 1076–1076, 1999Google Scholar
  38. 38.
    Mitchell SH, Zhu W, Young CYF: Resveratrol inhibits the expression and function of the androgen receptor in LNCaP prostate cancer cells. Cancer Res 59: 5892–5895, 1999Google Scholar
  39. 39.
    Israeli RS, Miller WHJ, Su SL, Samadi DS, Powell CT, Heston WDW, Wise GJ, Fair WR: Sensitive detection of prostatic hematogenous tumor cell dissemination using prostate specific antigen and prostate specific membrane-derived primers in the polymerase chain reaction. J Urol 153: 573–577, 1994Google Scholar
  40. 40.
    Dai JL, Maiorino CA, Gkonos PJ, Burnstein KL: Androgenic up-regulation of androgen receptor cDNA expression in androgen-independent prostate cancer cells. Steroids 61: 531–539, 1996Google Scholar
  41. 41.
    Fong CJ, Sherwood ER, Braun EJ, Berg LA, Lee C, Kozlowski JM: Regulation of prostatic carcinoma cell proliferation and secretory activity by extracellular matrix and stromal secretions. Prostate 21: 121–131, 1992Google Scholar
  42. 42.
    Guo Y, Pili R, Passaniti A: Regulation of prostate-specific antigen gene expression in LNCaP human prostatic carcinoma cells by growth, dihydrotestosterone, and extracellular matrix. Prostate 24: 1–10, 1994Google Scholar
  43. 43.
    Hsieh JT, Wu HC, Gleave ME, von Eschenbach AC, Chung LW: Autocrine regulation of prostate-specific antigen gene expression in a human prostatic cancer (LNCaP) subline. Cancer Res 53: 2852–2857, 1993Google Scholar
  44. 44.
    Wood CG, Sensibar J, Lee C, Kozlowski JM: Phenylacetate and phenylbutyrate promote cellular differentiation through downregulation of androgen receptor and bcl-2 levels in the prostate cancer cell line LNCaP (Meeting abstract). Proc Annu Meet Am Assoc Cancer Res 36: A3831, 1995Google Scholar
  45. 45.
    Wang LG, Liu XM, Kreis W, Budman DR: Down-regulation of prostate-specific antigen expression by finasteride through inhibition of complex formation between androgen receptor and steroid receptor-binding consensus in the promoter of the PSA gene in LNCaP cells, Cancer Res 57: 714–719, 1997Google Scholar
  46. 46.
    Thalmann GN, Sikes RA, Chang SM, Johnston DA, Von Eschenbach AC, Chung LWK: Suramin-induced decrease in prostate-specific antigen expression with no effect on tumor growth in the LNCaP model of human prostate cancer. Jnci 88: 794–801, 1996Google Scholar
  47. 47.
    Pettaway CA, Pathak S, Greene ME, Ramirez E, Wilson MR, Killion JJ, Fidler IJ: Selection of highly metastatic variants of different human prostatic carcinomas using orthotopic implantation in nude mice. Clin Cancer Res 2: 1627–1636, 1996Google Scholar
  48. 48.
    Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987Google Scholar
  49. 49.
    Israeli RS, Miller WH, Jr, Su SL, Powell CT, Fair WR, Samadi DS, Huryk RF, DeBlasio A, Edwards ET, Wise GJ et al: Sensitive nested reverse transcription polymerase chain reaction detection of circulating prostatic tumor cells: comparison of prostate-specific membrane antigen and prostate-specific antigen-based assays. Cancer Research 54: 6306–6310, 1994Google Scholar
  50. 50.
    Presnell SC, Borchert KM, Glover WJ, Gregory CW, Mohler JL, Smith GJ: Isolation and characterization of propagable cell lines (HUNC) from the androgen-sensitive Dunning R3327H rat prostatic adenocarcinoma. Carcinogenesis 19: 585–590, 1998Google Scholar
  51. 51.
    Peehl DM, Wong ST, Rubin JS: KGF and EGF differentially regulate the phenotype of prostatic epithelial cells. Growth Regul 6: 22–31, 1996Google Scholar
  52. 52.
    Young CY, Andrews PE, Tindall DJ: Expression and androgenic regulation of human prostate-specific kallikreins. J Androl 16: 97–99, 1995Google Scholar
  53. 53.
    Sadar MD: Androgen-independent induction of prostatespecific antigen gene expression via cross-talk between the androgen receptor and protein kinase A signal transduction pathways. J Biol Chem 274: 7777–7783, 1999Google Scholar
  54. 54.
    Wang LG, Liu XM, Kreis W, Budman DR: Androgen antagonistic effect of estramustine phosphate (EMP) metabolites on wild-type and mutated androgen receptor. Biochem Pharmacol 55: 1427–1433, 1998Google Scholar
  55. 55.
    Andrews PE, Young CY, Montgomery BT, Tindall DJ: Tumorpromoting phorbol ester down-regulates the androgen induction of prostate-specific antigen in a human prostatic adenocarcinoma cell line. Cancer Res 52: 1525–1529, 1992Google Scholar
  56. 56.
    Young CY, Montgomery BT, Andrews PE, Qui SD, Bilhartz DL, Tindall DJ: Hormonal regulation of prostate-specific antigen messenger RNA in human prostatic adenocarcinoma cell line LNCaP. Cancer Res 51: 3748–3752, 1991Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Barbara A. Woynarowska
    • 1
    • 2
  • Arlene L. Higdon
    • 1
    • 2
  • Ruben M. Muñoz
    • 1
    • 2
  • Perry Bushong
    • 1
  • Stephen J. Waters
    • 1
    • 2
  1. 1.Department of Radiation OncologyThe University of Texas Health Science CenterSan Antonio
  2. 2.MGI Pharma, Inc.BloomingtonU.S.A.

Personalised recommendations