Molecular and Cellular Biochemistry

, Volume 374, Issue 1–2, pp 49–59 | Cite as

Androgen deprivation by flutamide modulates uPAR, MMP-9 expressions, lipid profile, and oxidative stress: amelioration by daidzein

  • Abdul Lateef
  • Abdul Quaiyoom Khan
  • Mir Tahir
  • Rehan Khan
  • Muneeb U Rehman
  • Farrah Ali
  • Oday O. Hamiza
  • Sarwat Sultana


The growth and development of prostate gland is governed by testosterone. Testosterone helps in maintaining the adipose tissue stores of the body. It is well documented that with advancing age there has been a gradual decline in testosterone levels. Our aim was to study the protective role of daidzein on flutamide-induced androgen deprivation on matrix degrading genes, lipid profile and oxidative stress in Wistar rats. Sub-chronic (60 days) flutamide (30 mg/kg b.wt) administration resulted in marked increase in expressions of matrix degrading genes [matrix metalloproteases 9 and urokinase plasminogen activation receptor]. Additionally, it increased the levels of low density lipoproteins, total cholesterol, triglycerides, and lowered the levels of high density lipoproteins and endogenous antioxidant levels. Oral administration of daidzein (20 and 60 mg/kg b.wt) restituted the levels to normal. Daidzein administration resulted in amelioration of the prostate atrophy, degeneracy and invasiveness induced by flutamide. Our findings suggest that the daidzein may be given as dietary supplement to patients who are on androgen deprivation therapy, to minimize the adverse effects related to it and also retarding susceptibility of patients to cardiovascular diseases.


Androgen receptor (AR) Flutamide Matrix metalloproteases 9 (MMP-9) Daidzein Urokinase plasminogen activation receptor (uPAR) Cholesterol 



Androgen receptor


Benign prostatic hyperplasia


1-Chloro 2,4-dinitrobenzene


Cancer of prostate




Cardiovascular diseases


5,5′-Dithio-bis [2-nitrobenzoic acid]


Ethylenediamine tetra acetic acid


Glutathione reductase


Reduced glutathione


Oxidized glutathione


High density lipoprotein


Low density lipoprotein


Matrix metalloproteases 9


Post-mitochondrial supernatant


Reduced nicotinamide adenine dinucleotide phosphate


Reactive oxygen species




Urokinase plasminogen activation receptor


Ventral prostate



The author (SS) is thankful to the Department of Biotechnology (DBT), Govt. of India, New Delhi for providing funds and SRF to her student (AL) to carry out this work.


  1. 1.
    Raghow S, Kuliyev E, Steakley M, Greenberg N, Steiner MS (2000) Efficacious chemoprevention of primary prostate cancer by flutamide in an autochthonous transgenic model. Cancer Res 60:4093–4097PubMedGoogle Scholar
  2. 2.
    Higano CS (2003) Side effects of androgen deprivation therapy: monitoring and minimizing toxicity. Urology 61(2 Suppl 1):32–38PubMedCrossRefGoogle Scholar
  3. 3.
    Marin P, Holmang S, Jonsson L, Sjostrom L, Kvist H, Holm G, Lindstedt G, Björntorp P (1992) The effects of testosterone treatment on body composition and metabolism in middle-aged obese men. Int J Obes Relat Metab Disord 16:991–997PubMedGoogle Scholar
  4. 4.
    Satariano WA, Ragland K, Van Den Eeden S (1998) Cause of death in men diagnosed with prostate carcinoma. Cancer 83:1180–1188PubMedCrossRefGoogle Scholar
  5. 5.
    Lu-Yao G, Stukel TA, Yao SL (2004) Changing patterns in competing causes of death in men with prostate cancer: a population based study. J Urol 171:2285–2290PubMedCrossRefGoogle Scholar
  6. 6.
    Sprenkle P, Fisch H (2007) Pathologic effects of testosterone deprivation. Curr Opin Urol 17:424–430PubMedCrossRefGoogle Scholar
  7. 7.
    Saigal CS, Gore JL, Krupski TL, Hanley J, Schonlau M, Litwin MS (2007) Androgen deprivation therapy increases cardiovascular morbidity in men with prostate cancer. Cancer 110:1493–1500PubMedCrossRefGoogle Scholar
  8. 8.
    Zou Z, Zeng F, Xu W, Wang C, Ke Z, Wang QJ, Deng F (2012) PKD2 and PKD3 promote prostate cancer cell invasion via uPA by shifting balance between NF-κB and HDAC1. J Cell Sci. doi: 10.1242/jcs.106542
  9. 9.
    Kotipatruni RR, Nalla AK, Asuthkar S, Gondi CS, Dinh DH, Rao JS (2012) Apoptosis induced by knockdown of uPAR and MMP-9 is mediated by inactivation of EGFR/STAT3 signaling in medulloblastoma. PLoS ONE 7(9):e44798PubMedCrossRefGoogle Scholar
  10. 10.
    Jeffery N, McLean MH, El-Omar EM, Murray GI (2009) The matrix metalloproteinase/tissue inhibitor of matrix metalloproteinase profile incolorectal polyp cancers. Histopathology 54(7):820–828PubMedCrossRefGoogle Scholar
  11. 11.
    Nagase H, Woessner JF Jr (1999) Matrix metalloproteinases. J Biol Chem 274:21491–21494PubMedCrossRefGoogle Scholar
  12. 12.
    Bostwick DG, Alexander EE, Singh R, Shan A, Qian J, Santella RM, Oberley LW, Yan T, Zhong W, Jiang X, Oberley TD (2000) Antioxidantenzyme expression and reactive oxygen species damage in prostatic intraepithelial neoplasia and cancer. Cancer 89:123–134PubMedCrossRefGoogle Scholar
  13. 13.
    Oberley TD, Zhong W, Szweda LI, Oberley LW (2000) Localization of anti-oxidant enzymes and oxidative damage products in normal and malignant prostate epithelium. Prostate 44:144–155PubMedCrossRefGoogle Scholar
  14. 14.
    Masilamani M, Wei J, Sampson HA (2012) Regulation of the immune response by soybean isoflavones. Immunol Res 54(1–3):95–110Google Scholar
  15. 15.
    Messina M (2010) A brief historical overview of the past two decades of soy and isoflavone research. J Nutr 140:1350S–1354SPubMedCrossRefGoogle Scholar
  16. 16.
    Mishra P, Kar A, Kale RK (2009) Prevention of chemically induced mammary tumorigenesis by daidzein in pre-pubertal rats: the role of peroxidative damage and antioxidative enzymes. Mol Cell Biochem 325:149–157PubMedCrossRefGoogle Scholar
  17. 17.
    Wijeratne SS, Cuppett SL (2007) Soy isoflavones protect the intestine from lipid hydroperoxide mediated oxidative damage. J Agric Food Chem 55:9811–9816PubMedCrossRefGoogle Scholar
  18. 18.
    Jackman KA, Woodman OL, Sobey CG (2007) Isoflavones, equol and cardiovascular disease: pharmacological and therapeutic insights. Curr Med Chem 14:2824–2830PubMedCrossRefGoogle Scholar
  19. 19.
    Torres N, Torre-Villalvazo I, Tovar AR (2006) Regulation of lipid metabolism by soy protein and its implication in diseases mediated by lipid disorders. J Nutr Biochem 17:365–373PubMedCrossRefGoogle Scholar
  20. 20.
    Wright JR, Colby HD, Miles PR (1981) Cytosolic factors which affect microsomal lipid peroxidation in lung and liver. Arch Biochem Biophys 206:296–304PubMedCrossRefGoogle Scholar
  21. 21.
    Jollow DJ, Mitchell JR, Zampaglione N, Gillette JR (1974) Bromobenzene-induced liver necrosis. protective role of glutathione and evidence for 3,4-bromobenzene oxide as the hepatotoxic metabolite. Pharmacology 11:151–169PubMedCrossRefGoogle Scholar
  22. 22.
    Claiborne A (1985) Catalase activity. In: Greenwald RA (ed) CRC handbook of methods in oxygen radical research. CRC, Boca Raton, pp 283–284Google Scholar
  23. 23.
    Mohandas M, Marshall JJ, Duggin GG, Horvath JS, Tiller D (1984) Differential distribution of glutathione and glutathione-related enzymes in rabbit kidney: possible implications in analgesic nephropathy. Biochem Pharmacol 33:1801–1807PubMedCrossRefGoogle Scholar
  24. 24.
    Carlberg I, Mannervik B (1975) Glutathione level in rat brain. J Biol Chem 250:5475–5480PubMedGoogle Scholar
  25. 25.
    Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR (1982) Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Anal Biochem 126:131–138PubMedCrossRefGoogle Scholar
  26. 26.
    Marklund S, Marklund G (1974) Involvement of the superoxide anion in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474PubMedCrossRefGoogle Scholar
  27. 27.
    Michael Pfaffl W (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:2003–2007CrossRefGoogle Scholar
  28. 28.
    Vaibhav K, Shrivastava P, Javed H, Khan A, Ahmed HE, Tabassum R, Khan MM, Khuwaja G, Islam F, Saeed Siddiqui M, Safhi MM, Islam F (2012) Piperine suppresses cerebral ischemia reperfusion-induced inflammation through the repression of COX-2, NOS-2, and NF-κB in middle cerebral artery occlusion rat model. Mol Cell Biochem 367:73–84PubMedCrossRefGoogle Scholar
  29. 29.
    Lowry OH, Rosebrough NJ, Farr A, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  30. 30.
    Zgliczynski S, Ossowski M, Slowinska-Srzednicka J, Brzezinska A, Zgliczynski W, Soszynski P, Chotkowska E, Srzednicki M, Sadowski Z (1996) Effect of testosterone replacement therapy on lipids and lipoproteins in hypogonadal and elderly men. Atherosclerosis 121:35–43PubMedCrossRefGoogle Scholar
  31. 31.
    Braga-Basaria M, Muller D, Carducci MA, Dobs A, Basaria S (2006) Lipoprotein profile in men with prostate cancer undergoing androgen deprivation therapy. Int J Impot Res 18:494–498PubMedCrossRefGoogle Scholar
  32. 32.
    Smith MR, Nathan DM, Lee HW (2006) Insulin sensitivity during combined androgen blockade for prostate cancer. J Clin Endocrinol Metab 91:1305–1308PubMedCrossRefGoogle Scholar
  33. 33.
    Yannucci J, Manola J, Garnick MB, Bhat G, Bubley G (2006) The effect of androgen deprivation therapy on fasting serum lipid and glucose parameters. J Urol 176:520–525PubMedCrossRefGoogle Scholar
  34. 34.
    Chen KC, Peng CC, Hsieh HM, Peng CH, Hsieh CL, Huang CN, Chyau CC, Wang HE, Peng RY (2005) Antiandrogenic therapy can cause coronary arterial disease. Int J Urol 12:886–891PubMedCrossRefGoogle Scholar
  35. 35.
    Prins GS, Birch L (1995) The developmental pattern of androgen receptor expression in rat prostate lobes is altered after neonatal exposure to estrogen. Endocrinology 136(3):1303–1314PubMedCrossRefGoogle Scholar
  36. 36.
    Ishioka J, Hara S, Isaacs JT, Tomura A, Nishikawa K, Kageyama Y (2009) Suppression of mutant androgen receptors by flutamide. Int J Urol 16(5):516–521PubMedCrossRefGoogle Scholar
  37. 37.
    Dass K, Ahmad A, Azmi AS, Sarkar SH, Sarkar FH (2008) Evolving role of uPA/uPAR system in human cancers. Cancer Treat Rev 34:122–136PubMedCrossRefGoogle Scholar
  38. 38.
    Bugge TH, Lund LR, Kombrinck KK, Nielsen BS, Holmback K, Drew AF, Rick MJ, Witte DP, Dane K, Degen JL (1998) Reduced metastasis of mammary cancer in plasminogen-deficient mice. Oncogene 16:3097–3104PubMedCrossRefGoogle Scholar
  39. 39.
    Dane K, GrendahI-Hansen J, Eriksen J, Nielsen BS, Remer J, Pyke C (1993) The receptor for urokinase plasminogen activator. Stromal cell involvement in extracellular proteolysis during cancer invasion. In: Barrett AJ, Bond J (eds) Proteolysis and protein turnover. Portland Press, London, pp 239–245Google Scholar
  40. 40.
    Mc Carthy SM, Peter BF, Matthews DE, Akaike T, van der Vliet A (2008) Nitric oxide regulation of MMP-9 activation and its relationship to modifications of the cysteine switch. Biochemistry 47:5832–5840CrossRefGoogle Scholar
  41. 41.
    Benassayag C, Perrot-Applanat M, Ferre F (2002) Phytoestrogens as modulators of steroid action in target cells. J Chromatogr 777:233–248CrossRefGoogle Scholar
  42. 42.
    Dulak J, Józkowicz A, Dembinska-Kiec A, Guevara I, Zdzienicka A, Zmudzinska-Grochot D, Florek I, Wójtowicz A, Szuba A, Cooke JP (2000) Nitric oxide induces the synthesis of vascular endothelial growth factor by rat vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 20:659–666PubMedCrossRefGoogle Scholar
  43. 43.
    Ziche M, Parenti A, Ledda F, Dell’Era P, Granger HJ, Maggi CA, Presta M (1997) Nitric oxide promotes proliferation and plasminogen activator production by coronary venular endothelium through endogenous bFGF. Circ Res 80:845–852PubMedCrossRefGoogle Scholar
  44. 44.
    Joussen AM, Rohrschneider K, Reichling J, Kirchhof B, Kruse FE (2000) Treatment of corneal neovascularization with dietary isoflavonoids and flavonoids. Exp Eye Res 71:483–487PubMedCrossRefGoogle Scholar
  45. 45.
    Nohl H, Kozlov AV, Gille L, Staniek K (2003) Cell respiration and formation of reactive oxygen species: facts and artifacts. Biochem Soc Trans 31:1308–1311PubMedCrossRefGoogle Scholar
  46. 46.
    Mates JM, Perez-Gomez C, Nunez DCI (1999) Antioxidant enzymes and human diseases. Clin Biochem 32:595–603PubMedCrossRefGoogle Scholar
  47. 47.
    Dickinson DA, Forman HJ (2002) Glutathione in defense and signaling: lessons from a small thiol. Ann NY Acad Sci 973:488–504PubMedCrossRefGoogle Scholar
  48. 48.
    Shiota M, Yokomizo A, Tada Y, Inokuchi J, Kashiwagi E, Masubuchi D, Eto E, Uchiumi T, Naito S (2009) Castration resistance of prostate cancer cells caused by castration-induced oxidative stress through Twist1 and androgen receptor overexpression. Oncogene 29:237–250PubMedCrossRefGoogle Scholar
  49. 49.
    Shiota M, Yokomizo A, Naito S (2011) Oxidative stress and androgen receptor signaling in the development and progression of castration-resistant prostate cancer. Free Radic Biol Med 51:1320–1328PubMedCrossRefGoogle Scholar
  50. 50.
    Sichel G, Corsaro C, Scalia M, Di Bilio AJ, Bonomo RP (1991) In vitro scavenger activity of some flavonoids and melanins against O2−. Free Radic Biol Med 11:1–8PubMedCrossRefGoogle Scholar
  51. 51.
    Hanashi Y, Ogawa S, Fukui S (1994) The correlation between active oxygen scavenging and antioxidative effects of flavonoids. Free Radic Biol Med 16:845–850CrossRefGoogle Scholar
  52. 52.
    Rohrdanz E, Ohler S, Tran-Thi QH, Kahl R (2002) The phytoestrogen daidzein affects the antioxidant enzyme system of rat hepatoma H4IIE cells. J Nutr 132:370–375PubMedGoogle Scholar
  53. 53.
    Marnett LJ (2000) Oxyradicals and DNA damage. Carcinogenesis 21:361–370PubMedCrossRefGoogle Scholar
  54. 54.
    Parola M, Belloma G, Robino G, Barrera G, Dianzani MU (1999) 4-Hydroxynonenal as a biological signal: molecular basis and pathophysiological implications. Antioxid Redox Signal 1:255–284PubMedCrossRefGoogle Scholar
  55. 55.
    Winrow VR, Winyard PG, Morris CJ, Black DR (1993) Free radicals in inflammation: second messengers and mediators of tissue destruction. Br Med Bull 49:506–522PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Abdul Lateef
    • 1
  • Abdul Quaiyoom Khan
    • 1
  • Mir Tahir
    • 1
  • Rehan Khan
    • 1
  • Muneeb U Rehman
    • 1
  • Farrah Ali
    • 1
  • Oday O. Hamiza
    • 1
  • Sarwat Sultana
    • 1
  1. 1.Molecular Carcinogenesis and Chemoprevention Division, Department of Medical Elementology and Toxicology, Faculty of ScienceJamia Hamdard (Hamdard University)New DelhiIndia

Personalised recommendations