Skip to main content

Advertisement

Log in

Effects of diethylstilbestrol in human lymphocytes in vitro: A dose and time-dependent study on genotoxic, cytotoxic and apoptotic effects

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Most of the biological, chemical or physical agents that cause cell death in certain doses and time of exposure may induce either apoptosis or necrosis. This study explores in what ways the genotoxic, cytotoxic and apoptotic effects of diethylstilbestrol (DES), a chemical agent currently used in the treatment of various types of cancer, on the human lymphocytes depend upon the dose and the exposure time. For this purpose, firstly it aims to determine in what dosages and durations of DES treatment, genotoxicity and cytotoxicity in human lymphocytes occur in vitro. Secondly, it explores the effects of DES on sister-chromatid exchanges (SCEs) and apoptosis and their relation with the nitric oxide (NO) levels. Finally, it investigates whether different dosages of DES and duration of treatment with it are correlated with each other. In so doing, we investigated the relationship among the viability, necrosis and apoptosis rates of human lymphocytes which were treated with five different DES concentrations (1, 5, 10, 15 and 20 μM) for 24, 48 and 72 h, DNA fragmentation analysis of these cells, their mean SCE values and NO levels. We concluded that 5 μM DES at 24 h is the most effective dosage that induces typical features of apoptosis in human lymphocytes. Despite the fact that there are many other studies on the effects of DES on the cancer cells, we thought it might be worth looking into the effects of DES on human lymphocytes in vitro. We meant the present study to contribute to the research done in the field of cancer treatment. (Mol Cell Biochem 276: 45–53, 2005)

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

AO/EtBr:

acridine orange/ethidium bromide

DES:

diethylstilbestrol

NO:

nitric oxide

SCEs:

sister-chromatid exchanges

References

  1. Bennink Coelingh HJT: Are all estrogens the same? Maturitas 47: 269–275, 2004

    Google Scholar 

  2. Marselos M, Tomatis L: Diethylstilbestrol: I. Pharmacology, toxicology and carcinogenicity in humans. Eur J Cancer 28A: 1182–1189, 1992

    Article  PubMed  Google Scholar 

  3. Herbst AL, Ulfelder H, Poskanzer DC: Adenocarcinoma of the vagina: Association of the maternal stilbestrol therapy with tumor appearance in young women. New Engl J Med 284: 878–881, 1971

    PubMed  Google Scholar 

  4. Degen GH, Bolt HM: Endocrine disruptors: Update on xenoestrogens. Int Arch Occup Environ Health 73: 433–441, 2000

    Article  PubMed  Google Scholar 

  5. Banduhn N, Obe G: Mutagenicity of methyl 2-benzimidazolecarbamate, diethylstilbestrol and estradiol: Structural chromosomal aberrations, sister-chromatid exchanges, C-mitoses, polyploidies and micronuclei. Mutat Res-Gen Toxicol 156(3): 199–218, 1985

    Article  Google Scholar 

  6. Sharma RK, Jacobson-Kram D, Lemmon M, Bakke J, Galperin I, Blazak WF: Sister-chromatid exchange and cell replication kinetics in fetal and maternal cells after treatment with chemical teratogens. Mutat Res-Gen Toxicol 158(3): 217–231, 1985

    Article  Google Scholar 

  7. Schuler M, Hasegawa L, Parks R, Metzler M, Eastmond DA: Dose-response studies of the induction of hyperdiploidy and polyploidy by diethylstilbestrol and 17 beta-estradiol in cultured human lymphocytes using multicolor fluorescence in situ hybridization. Environ Mol Mutagen 31(3): 263–73, 1998

    Article  PubMed  Google Scholar 

  8. Cox R, Crawford E: Estrogens in the treatment of prostate cancer. J Urol 154: 1991–1998, 1995

    Article  PubMed  Google Scholar 

  9. Calemine JB, Gogal RM, Lengi A, Sponenberg P, Ahmed SA: Immunomodulation by diethylstilbestrol is dose and gender related: effects on thymocyte apoptosis and mitogen-induced proliferation. Toxicol 178: 101–118, 2002

    Article  Google Scholar 

  10. Scherr D, Pitts WR, Vaughan ED: Diethylstilbestrol revisited: Androgen deprivation, osteoporosis and prostate cancer. J Urol 167: 535–538, 2002

    Article  PubMed  Google Scholar 

  11. Kerr JF, Wyllie AH, Currie AR: Apoptosis: A basic biological phenomenon with wide-ranging implication in tissue kinetics. Br J Cancer 26: 239–257, 1972

    PubMed  Google Scholar 

  12. Robertson CN, Roberson KM, Padillla GM, O’Brien ET, Cook JM, Kim CS, Fine RL: Induction of apoptosis by diethylstilbestrol in hormone-insensitive prostate cancer cells. J Natl Cancer Inst 88(13): 908–917, 1996

    PubMed  Google Scholar 

  13. Guo C, Yu S, Davis AT, Wang H, Green JE, Ahmed K: A potential role of nuclear matrix-associated protein kinase CK2 in protection against drug-induced apoptosis in cancer cells. J Biol Chem 276(8): 5992–5999, 2001

    Article  PubMed  Google Scholar 

  14. Qui CH, Ohe M, Koibuchi N, Matsuzaki S: Apoptosis in the epididymal epithelium of adult male golden hamster exposed to diethylstilbestrol. J Histochem Cytochem 52(2): 187–192, 2004

    PubMed  Google Scholar 

  15. Bursch W, Liehr JG, Sirbasku DA, Putz B, Taper H, Schulte-Hermann R: Control of cell Death (apoptosis) by diethylstilbestrol in an estrogen-dependent kidney tumor. Carcinogenesis 12(5): 855–860, 1991

    PubMed  Google Scholar 

  16. Nair R, Shaha C: Diethylstilbestrol Induces Rat Spermatogenic Cell Apoptosis in vivo through increased expression of spermatogenic cell Fas/FasL system. J Biol Chem 278(8): 6470–6481, 2003

    Article  PubMed  Google Scholar 

  17. Lai ZW, Fiore CN, Hahn PJ, Gasiewicz TA, Silverstone AE: Differential effects of diethylstilbestrol and 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin on thymocyte differentiation, proliferation and apoptosis in bcl-2 trangenic mouse fetal thymus organ culture. Toxicol Appl Pharm 168: 15–24, 2000

    Article  Google Scholar 

  18. Tomimori Y, Ikawa Y, Oyaizu N: Ultraviolet-irradiated apoptotic lymphocytes produce interleukin-10 by themselves. Immunol Lett 71(1): 49–54, 2000

    Article  PubMed  Google Scholar 

  19. Devadas S, Hinshaw JA, Zaritskaya L, Williams MS: Fas-stimulated generation of reactive oxygen species or exogenous oxidative stress sensitize cells to Fas-mediated apoptosis. Free Radic Biol Med 35(6): 648–661, 2003

    Article  PubMed  Google Scholar 

  20. Buzard GS, Kasprzak KS: Possible roles nitric oxide and redox cell signaling in toxicity and carcinogenesis: a review. J Environ Pathol Toxicol Oncol 19: 179–199, 2000

    PubMed  Google Scholar 

  21. Roger R, Issaad C, Pallardy M, Leglise MC, Turhan AG, Bertoglio J, Breard J: BCR-ABL does not prevent apoptotic death induced by human natural killer or lymphocyte-activated killer cells. Blood 87: 1113–1117, 1996

    PubMed  Google Scholar 

  22. Gong JP, Traganos F, Darzynkiewicz Z: A selective procedure for DNA extraction from apoptotic cells applicable for gel electrophoresis and flow cytometry. Anal Biochem 218(2): 314–319, 1994

    Article  PubMed  Google Scholar 

  23. Perry P, Wolff S: New Giemsa method for the differential staining of sister chromatids. Nature 258: 156–158, 1977

    Google Scholar 

  24. Braman RS, Hendrix SA: Nanogram nitrite and nitrate determination in environmental and biological materials by vanadium (III) reduction with chemiluminescence detection. Anal Chem 61: 2715–2718, 1989

    Article  PubMed  Google Scholar 

  25. Bratton SB, Cohen GM: Apoptotic death sensor: an organelle’s alter ego? Trends Pharmacol Sci 22: 306–315, 2001

    Article  PubMed  Google Scholar 

  26. Clarke AR, Purdie CA, Harrison DJ, Morris RG, Bird CC, Hooper ML, Wyllie AH: Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature 362: 849–852, 1993

    Article  PubMed  Google Scholar 

  27. Ritke MK, Rusnak JM, Lazo JS, Allan WP, Dive C, Heer S, Yalowich JC: Differential induction of etoposide-mediated apoptosis in human leukemia HL-60 and K-562 cells. Mol Pharmacol 46: 605–611, 1994

    PubMed  Google Scholar 

  28. Ergun MA, Konac E, Erbas D, Ekmekci A: Apoptosis and nitric oxide release induced by thalidomide, gossypol and dexamethasone in cultured human chronic myelogenous leukemic K-562 cells. Cell Biol Int 28: 237–242, 2004

    Article  PubMed  Google Scholar 

  29. Yang J, Duerksen-Hughes P: A new approach to identifying genotoxic carcinogens: p53 induction as an indicator of genotoxic damage. Carcinogenesis 19(6): 1117–1125, 1998

    Article  PubMed  Google Scholar 

  30. Smith DC, Redman BG, Flaherty LE, Li L, Strawderman M, Pienta KJ.: A Phase II trial of oral diethylstilbestrol as a second-line hormonal agent in advanced prostate cancer. Urology 52: 257–260, 1998

    Article  PubMed  Google Scholar 

  31. Ahmed M, Choksy S, Chilton CP, Munson KW, Williams JH: High dose intravenous oestrogen (fosfestrol) in the treatment of symptomatic, metastatic, hormone-refractory carcinoma of the prostate. Int Urol Nephrol 30: 159–164, 1998

    PubMed  Google Scholar 

  32. de Voogt HJ, Smith PH, Pavone-Macaluso M, de Pauw M, Suciu S: Cardiovascular side effects of diethylstilbestrol, cyproterone acetate, medroxyprogesterone acetate and estramustine phosphate used for the treatment of advanced prostatic cancer: Results from European Organization for Research on Treatment of Cancer Trials 30761 and 30762. J Urol 135: 303–307, 1986

    PubMed  Google Scholar 

  33. Ham JM, Pirola RC, Crouch RL: Hemangioendothelial sarcoma of the liver associated with long-term estrogen therapy in a man. Dig Dis Sci 25: 879–883, 1980

    Article  PubMed  Google Scholar 

  34. Brooks JJ: Hepatoma associated with diethylstilbestrol therapy for prostatic carcinoma. J Urol 128: 1044–1045, 1982

    PubMed  Google Scholar 

  35. Carmichael PL, Mills JJ, Campbell M, Basu M, Caldwell J: Mechanisms of hormonal carcinogenesis in the p53+/− hemizygous knockout mouse: Studies with diethylstilbestrol. Toxicol Pathol 29: 155–160, 2001

    Article  PubMed  Google Scholar 

  36. Marselos M, Tomatis L: Diethylstilbestrol II. Pharmacology, toxicology and carcinogenicity in humans. Eur J Cancer 29: 1182–1189, 1993

    Article  Google Scholar 

  37. Fluckiger-Isler S, Baumeister M, Braun K, Gervais V, Hasler-Nguyen N, Reimann R, Van Gompel J, Wunderlich HG, Engelhardt G: Assessment of the performance of the Ames IITMassay: A collaborative study with 19 coded compounds. Mutat Res-Gen Toxicol Eng 558(1–2): 181–197, 2004

    Article  Google Scholar 

  38. Dreher D, Junod AF: Role of oxygen free radicals in cancer development. Eur J Cancer 32A(1): 30–38, 1996

    Article  PubMed  Google Scholar 

  39. Igney FH, Krammer PH: Death and anti-death: Tumor resistance to apoptosis. Nature 2: 277–288, 2002

    Google Scholar 

  40. Kolb JP: Mechanisms involved in the pro- and anti-apoptotic effects of NO in human leukemia. Leukemia 14: 1685–1694, 2000

    Article  PubMed  Google Scholar 

  41. Zhang WY, Gotoh T, Oyadomaria S, Mori M: Coinduction of inducible nitric oxide synthase and arginine recycling enzymes in cytokine-stimulated PC12 cells and high output production of nitric oxide. Mol Brain Res 83: 1–8, 2000

    Article  PubMed  Google Scholar 

  42. Mannick JB, Miao XQ, Stamler JS: Nitric oxide inhibits Fas-induced apoptosis. J Biol Chem 272(39): 24125–24128, 1997

    Article  PubMed  Google Scholar 

  43. Sciorati C, Rovere P, Ferrarini M, Heltai S, Manfredi AA, Clementi E: Autocrine nitric oxide modulates CD95-induced apoptosis in gammadelta T lymphocytes. J Biol Chem 272(37): 23211–23215, 1997

    Article  PubMed  Google Scholar 

  44. Albina JE, Cui S, Mateo RB, Reichner JS: Nitric oxide-mediated apoptosis in murine peritoneal macrophages. J Immunol 150: 5080–5085, 1993

    PubMed  Google Scholar 

  45. Kaneto H, Fujii J, Seo HG, Suzuki K, Matsuoka T, Nakamura M, Tatsumi H, Yamasaki Y, Kamada T, Taniguchi N: Apoptotic cell death triggered by nitric oxide in pancreatic beta-cells. Diabetes 44(7): 733–738, 1995

    PubMed  Google Scholar 

  46. Valenti L, Mathieu J, Chancerelle Y, Levacher M, Chanaud B, De Sousa M, Strzalko S, Dinh-Xuan AT, Giroud JP, Florentin I: Nitric oxide inhibits spleen cell proliferative response after burn injury by inducing cytostasis, apoptosis and necrosis of activated T lymphocytes: Role of the guanylate cyclase. Cell Immunol 221: 50–63, 2003

    Article  PubMed  Google Scholar 

  47. Brune B, Von Knethen A, Sandau KB: Nitric oxide and its role in apoptosis. Eur J Pharmacol 351: 172–261, 1998

    Google Scholar 

  48. Joshi MS, Ponthier JL, Lancaster JR: Cellular antioxidant and pro-oxidant actions of nitric oxide. Free Radical Bio Med 27: 1357–1366, 1999

    Article  Google Scholar 

  49. Bonfoco E, Krainc D, Ankarcron M, Nicotera P, Lipton SA: Apoptosis and necrosis: Two distinct events induced, respectively, by mild and intense insults with N-methyl-D-aspartate or nitric oxide/superoxide in cortical cell cultures. Proc Natl Acad Sci 92: 7162–7166, 1995

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ece Konac.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Konac, E., Ekmekci, A., Barkar, V. et al. Effects of diethylstilbestrol in human lymphocytes in vitro: A dose and time-dependent study on genotoxic, cytotoxic and apoptotic effects. Mol Cell Biochem 276, 45–53 (2005). https://doi.org/10.1007/s11010-005-2815-8

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-005-2815-8

Keywords

Navigation