Czech J. Anim. Sci., 2009, 54(12):566-574 | DOI: 10.17221/136/2009-CJAS

Effect of aromatase inhibitor (fadrozole) on proliferation, estradiol production and telomerase activity in pig granulosa cells in vitro

E. Chronowska1, M. Tománek1, T. Kott2
1 Institute of Animal Science, Department of Biology of Reproduction, Prague-Uhříněves, Czech Republic
2 Institute of Animal Science, Department of Molecular Genetics, Prague-Uhříněves, Czech Republic

The objective of the present work was to study the effect of a nonsteroidal aromatase inhibitor (fadrozole) on proliferation, estradiol production, aromatase expression and telomerase activity (TA) in pig granulosa cells (GC) from small (1-2 mm) and large (5-7 mm) follicles. The cells were treated with fadrozole for 48 h and 72 h in basal and FSH-stimulated conditions. Fadrozole caused a decrease (P < 0.05) of 3H-thymidine incorporation in granulosa cells derived from small (1-2 mm) and large follicles (5-7 mm). The proliferative potential of small-follicle GC was significantly higher (P < 0.01) under all culture conditions. Estradiol production was suppressed (P < 0.01) in both granulosa cell populations cultured in the presence of fadrozole for 48 and 72 h. Fadrozole caused a decrease (P < 0.05) of aromatase gene expression in small-follicle granulosa cell incubated for 72 h and in large-follicle GC after 48 h of culture. Large-follicle GC were characterized by a higher (P < 0.01) level of estradiol production and aromatase gene expression. Telomerase activity decreased (P < 0.05) in large-follicle granulosa cells incubated in the presence of an aromatase inhibitor for 72 h. The TA level in large-follicle granulosa cells was higher (P < 0.01) in comparison to small-follicle GC in all culture conditions after 72 h of incubation. The results of the present study suggest the important role of telomerase in the process of follicular growth and development.

Keywords: granulosa cells; aromatase inhibitor; proliferation; telomerase; estradiol

Published: December 31, 2009  Show citation

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Chronowska E, Tománek M, Kott T. Effect of aromatase inhibitor (fadrozole) on proliferation, estradiol production and telomerase activity in pig granulosa cells in vitro. Czech J. Anim. Sci. 2009;54(12):566-574. doi: 10.17221/136/2009-CJAS.
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References

  1. Bayne S., Liu J.P. (2005): Hormones and growth factors regulate telomerase activity in ageing and cancer. Molecular and Cellular Endocrinology, 240, 11-22. Go to original source... Go to PubMed...
  2. Bendell J.J., Dorrington J. (1991): Estradiol-17-beta stimulates DNA-synthesis in rat granulosa cells action mediated by transforming growth factor beta. Endocrinology, 128, 2663-2665. Go to original source... Go to PubMed...
  3. Billig H., Furuta I., Hsueh A.J. (1993): Estrogens inhibit and androgens enhance ovarian granulosa cell apoptosis. Endocrinology, 133, 2204-2212. Go to original source... Go to PubMed...
  4. Bley M.A., Saragueta P.E., Baranao J.L. (1997): Concerted stimulation of rat granulosa cell deoxyribonucleic acid synthesis by sex steroids and follicle-stimulating hormone. Journal of Steroid Biochemistry and Molecular Biology, 62, 11-19. Go to original source... Go to PubMed...
  5. Brunner N., Bronzert D., Vindelov L.L., Rygaard K., Spang-Thomsen M., Lippman M.E. (1989): Effect on growth and cell cycle kinetics of estradiol and tamofixen on MCF-7 human breast cancer cells grown in vitro and in nude mice. Cancer Research, 49, 1515-1520.
  6. Conley A., Corbin J., Smith T., Hinshelwood M., Liu Z., Simpson E. (1997): Porcine aromatases: studies on tissue-specific, functionally distinct izozymes from a single gene? Journal of Steroid Biochemistry and Molecular Biology, 61, 407-413. Go to original source...
  7. Drummond A., Findlay J. (1999): The role of estrogen in folliculogenesis. Molecular and Cellular Endocrinology, 151, 57-64. Go to original source... Go to PubMed...
  8. Escamilla-Hernandez R., Little-Ihrig L., Orwig K.E., Yoe J.M., Chandran U., Zeleznik A.J. (2008): Constitutively active protein kinase a qualitatively mimics the effects of follicle-stimulating hormone on granulosa cell differentiation. Molecular Endocrinology, 22, 1842-1852 Go to original source... Go to PubMed...
  9. Fitzpatrick S.L., Richards J.S. (1991): Regulation of cytochrome P450 aramatase messenger ribonucleic acid and activity by steroids and gonadotropins in rat granulosa cells. Endocrinology, 129, 1452-1462. Go to original source... Go to PubMed...
  10. Greider C.V. (1998): Telomerase activity, cell proliferation, and cancer. The Proceedings of the National Academy of Science of the United States of America, 6, 95, 90-92. Go to original source... Go to PubMed...
  11. Gregoraszczuk E.L. (1996): Large and small cells of the porcine corpus luteum: Differential capacity to secrete estradiol and aromatize exogenous androgen during mid-and late phase. Experimental and Clinical Endocrinology and Diabetes, 104, 278-283. Go to original source... Go to PubMed...
  12. Gregoraszczuk E.L., Oblonczyk K. (1996): Effect of specific aromatase inhibitor on oestradiol secretion by porcine corpora lutea at various stages of the luteal phase. Reproduction Nutrition Development, 36, 65-72. Go to original source... Go to PubMed...
  13. Hickey T.E., Marrocco D.L., Ritter L.J., Norman R.J., Gilchrist R.B., Armstrong D.T. (2005): Androgens augment the mitotic effects of oocyte-secreted factors and growth differentiation factor 9 on porcine granulosa cells. Biology of Reproduction, 73, 825-832. Go to original source... Go to PubMed...
  14. Hirshfield A.N. (1991): Development of follicles in the mammalian ovary. International Review of Cytology, 124, 43-101. Go to original source... Go to PubMed...
  15. Kim N.W., Piatyszek M.A., Prowse K.R., Harley C.B., West M.D., Ho P.L., Coviello G.M., Wright W.E., Weinrich S.L., Shay J.W. (1994): Specific association of human telomerase activity with immortal cells and cancer. Science, 266, 2011-2015. Go to original source... Go to PubMed...
  16. Kyo S., Takakura M., Kohama T., Inoue M. (1997): Telomerase activity in human endometrium. Cancer Research, 57, 610-614.
  17. Langhout D.J., Spicer L.J., Geisert R.D. (1991): Development of a culture system for bovine granulosa cells effects of growth hormone, estradiol and gonadotropins on cell proliferation, steroidogenesis and protein synthesis. Journal of Animal Science, 69, 3321-3334. Go to original source... Go to PubMed...
  18. Lavranos T.C., Mathis J.M., Latam S.E., Kalionis B., Shay J.W., Rodgers R.J. (1999): Evidence for ovarian granulosa stem cells: Telomerase activity and localization of the telomerase ribonucleic acid component in bovine ovarian follicles. Biology of Reproduction, 61, 358-366. Go to original source... Go to PubMed...
  19. Miller W.R. (2003): Aromatase inhibitors: Mechanism of action and role in the treatment of breast cancers. Seminars in Oncology, 30, 3-11. Go to original source... Go to PubMed...
  20. Misiti S., Nanni S., Fontemaggi G., Cong Y.S., Wen J.P., Hirte H.W., Piaggio G., Sacchi A., Pontecorvi A., Bacchetti S., Farsetki A. (2000): Induction of hTERT Expression and telomerase activity by estrogens in human ovary epithelium cells. Molecular and Cellular Biology, 20, 3764-3771. Go to original source... Go to PubMed...
  21. Petr J., Tománek M., Fulka J. Jr., Míka J., Jílek F. (1991): Effects of GnRH on preovulatory endocrinology and oocyte maturation in PMSG-superovulated cows. Animal Reproduction Science, 24, 37-52. Go to original source...
  22. Picton H.M., Campbell B.K., Hunter M.G. (1999): Maintenence of oestradiol production and expression of cytochrome P450 aromatase enzyme mRNA in longterm serum-free cultures of pig granulosa cells. Journal of Reproduction and Fertility, 115, 67-77. Go to original source... Go to PubMed...
  23. Ranson E.J., Picton H.M., Hunter M.G. (1997): Effects of testosterone and oestradiol on (H-3)-thymidine incorporation by porcine granulosa and theca cells. Animal Reproduction Science, 47, 229-236. Go to original source... Go to PubMed...
  24. Richards J.S. (1975): Estradiol receptor content in rat granulosa cells during follicular development. Endocrinology, 97, 1174-1184. Go to original source... Go to PubMed...
  25. Tománek M., Chronowska E., Kott T., Czerneková V. (2008): Telomerase activity in pig granulosa cells proliferating and differentiating in vitro. Animal Reproduction Science, 104, 284-298. Go to original source... Go to PubMed...
  26. Vacková I., Engelová M., Marinov I., Tománek M. (2003): Cell cycle synchronization of porcine granulosa cells in G1 stage with mimosine. Animal Reproduction Science, 77, 235-245. Go to original source... Go to PubMed...
  27. Yamagata Y., Nakamura Y., Umayahara K., Harad A., Takayama H., Sugino N., Kato H. (2002): Changes in telomerase activity in experimentally induced atretic follicles of immature rats. Endocrine Journal, 49, 589-595. Go to original source... Go to PubMed...
  28. Yokoyama Y., Takahashi K., Morshita S., Hashimoto M., Niwa K., Tamaya T. (1998): Telomerase activity in the human endometrium throughout the menstrual cycle. Molecular Human Reproduction, 4, 173-177. Go to original source... Go to PubMed...
  29. Younus J., Vandenberg T.A. (2005): A practical overview of aromatase inhibitors in postmenopausal women with hormone receptor-positive breast cancer. Bulletin of Cancer, 92, 39-44.

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