Regulation of catechol O-methyltransferase expression in granulosa cells

a potential role for follicular arrest in polycystic ovary syndrome

Sana M. Salih, Mohammad Jamaluddin, Salama A. Salama, Amin A. Fadl, Manubai Nagamani, Ayman Al-Hendy

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Objective: To investigate the regulation of catechol O-methyltransferase (COMT) expression in granulosa cells and assess potential effects of 2-methoxyestradiol (2-ME2) and COMT inhibitors on granulosa cell steroidogenesis and proliferation. Design and Setting: Controlled experimental study in an academic research laboratory. Intervention(s): JC410 porcine and HGL5 human granulosa cell lines were used for in vitro experiments. Effects of 2-ME2 and COMT inhibitor treatment on DNA proliferation and steroidogenesis were assessed by using Hoechst dye and p450SCC-luciferase reporter assays. Effects of dihydrotestosterone (DHT), insulin, and all-trans retinoic acid (ATRA) on COMT messenger RNA expression were investigated by using COMTP1 promoter-luciferase reporter and Northern blot. Main Outcome Measure(s): Granulosa cell steroidogenesis and proliferation following COMP inhibitor and 2-ME2 treatment. Regulation of COMT expression with DHT, insulin, and ATRA. Result(s): 2-Methoxyestradiol had a dual effect on granulosa cell proliferation and p450SCC- luciferase activity; low doses were stimulatory and high doses were inhibitory. Catechol O-methyltransferase inhibitor was associated with up to a 65% increase in JC410 cell number and a maximal 5.6-fold increase in p450SCC-luciferase activity at 20 μmol/L. Dihydrotestosterone, insulin, and ATRA all induced a dose-dependent increase in COMTP1-luciferase transactivation, as well as up-regulated COMT messenger RNA expression in granulosa cells. Conclusion(s): Catechol O-methyltransferase expression in granulosa cells was up-regulated by insulin, DHT, and ATRA. Catechol O-methyltransferase product, 2-ME2, decreased, whereas COMT inhibitor increased granulosa cell proliferation and steroidogenesis. These data suggest that COMT overexpression with subsequent increased level of 2-ME2 may lead to ovulatory dysfunction.

Original languageEnglish (US)
Pages (from-to)1414-1421
Number of pages8
JournalFertility and Sterility
Volume89
Issue number5 SUPPL.
DOIs
StatePublished - May 2008

Fingerprint

Catechol O-Methyltransferase
Granulosa Cells
Polycystic Ovary Syndrome
Luciferases
Dihydrotestosterone
Tretinoin
Cell Proliferation
Insulin
Messenger RNA
Northern Blotting
Transcriptional Activation
Coloring Agents
Swine
Cell Count
Outcome Assessment (Health Care)
Cell Line
Catechol O-Methyltransferase Inhibitors
DNA

Keywords

  • anovulation
  • Catechol O-methyltransferase expression
  • granulosa cell steroidogenesis and proliferation
  • methoxyestrogen

ASJC Scopus subject areas

  • Obstetrics and Gynecology

Cite this

Regulation of catechol O-methyltransferase expression in granulosa cells : a potential role for follicular arrest in polycystic ovary syndrome. / Salih, Sana M.; Jamaluddin, Mohammad; Salama, Salama A.; Fadl, Amin A.; Nagamani, Manubai; Al-Hendy, Ayman.

In: Fertility and Sterility, Vol. 89, No. 5 SUPPL., 05.2008, p. 1414-1421.

Research output: Contribution to journalArticle

Salih, Sana M. ; Jamaluddin, Mohammad ; Salama, Salama A. ; Fadl, Amin A. ; Nagamani, Manubai ; Al-Hendy, Ayman. / Regulation of catechol O-methyltransferase expression in granulosa cells : a potential role for follicular arrest in polycystic ovary syndrome. In: Fertility and Sterility. 2008 ; Vol. 89, No. 5 SUPPL. pp. 1414-1421.
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abstract = "Objective: To investigate the regulation of catechol O-methyltransferase (COMT) expression in granulosa cells and assess potential effects of 2-methoxyestradiol (2-ME2) and COMT inhibitors on granulosa cell steroidogenesis and proliferation. Design and Setting: Controlled experimental study in an academic research laboratory. Intervention(s): JC410 porcine and HGL5 human granulosa cell lines were used for in vitro experiments. Effects of 2-ME2 and COMT inhibitor treatment on DNA proliferation and steroidogenesis were assessed by using Hoechst dye and p450SCC-luciferase reporter assays. Effects of dihydrotestosterone (DHT), insulin, and all-trans retinoic acid (ATRA) on COMT messenger RNA expression were investigated by using COMTP1 promoter-luciferase reporter and Northern blot. Main Outcome Measure(s): Granulosa cell steroidogenesis and proliferation following COMP inhibitor and 2-ME2 treatment. Regulation of COMT expression with DHT, insulin, and ATRA. Result(s): 2-Methoxyestradiol had a dual effect on granulosa cell proliferation and p450SCC- luciferase activity; low doses were stimulatory and high doses were inhibitory. Catechol O-methyltransferase inhibitor was associated with up to a 65{\%} increase in JC410 cell number and a maximal 5.6-fold increase in p450SCC-luciferase activity at 20 μmol/L. Dihydrotestosterone, insulin, and ATRA all induced a dose-dependent increase in COMTP1-luciferase transactivation, as well as up-regulated COMT messenger RNA expression in granulosa cells. Conclusion(s): Catechol O-methyltransferase expression in granulosa cells was up-regulated by insulin, DHT, and ATRA. Catechol O-methyltransferase product, 2-ME2, decreased, whereas COMT inhibitor increased granulosa cell proliferation and steroidogenesis. These data suggest that COMT overexpression with subsequent increased level of 2-ME2 may lead to ovulatory dysfunction.",
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T2 - a potential role for follicular arrest in polycystic ovary syndrome

AU - Salih, Sana M.

AU - Jamaluddin, Mohammad

AU - Salama, Salama A.

AU - Fadl, Amin A.

AU - Nagamani, Manubai

AU - Al-Hendy, Ayman

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N2 - Objective: To investigate the regulation of catechol O-methyltransferase (COMT) expression in granulosa cells and assess potential effects of 2-methoxyestradiol (2-ME2) and COMT inhibitors on granulosa cell steroidogenesis and proliferation. Design and Setting: Controlled experimental study in an academic research laboratory. Intervention(s): JC410 porcine and HGL5 human granulosa cell lines were used for in vitro experiments. Effects of 2-ME2 and COMT inhibitor treatment on DNA proliferation and steroidogenesis were assessed by using Hoechst dye and p450SCC-luciferase reporter assays. Effects of dihydrotestosterone (DHT), insulin, and all-trans retinoic acid (ATRA) on COMT messenger RNA expression were investigated by using COMTP1 promoter-luciferase reporter and Northern blot. Main Outcome Measure(s): Granulosa cell steroidogenesis and proliferation following COMP inhibitor and 2-ME2 treatment. Regulation of COMT expression with DHT, insulin, and ATRA. Result(s): 2-Methoxyestradiol had a dual effect on granulosa cell proliferation and p450SCC- luciferase activity; low doses were stimulatory and high doses were inhibitory. Catechol O-methyltransferase inhibitor was associated with up to a 65% increase in JC410 cell number and a maximal 5.6-fold increase in p450SCC-luciferase activity at 20 μmol/L. Dihydrotestosterone, insulin, and ATRA all induced a dose-dependent increase in COMTP1-luciferase transactivation, as well as up-regulated COMT messenger RNA expression in granulosa cells. Conclusion(s): Catechol O-methyltransferase expression in granulosa cells was up-regulated by insulin, DHT, and ATRA. Catechol O-methyltransferase product, 2-ME2, decreased, whereas COMT inhibitor increased granulosa cell proliferation and steroidogenesis. These data suggest that COMT overexpression with subsequent increased level of 2-ME2 may lead to ovulatory dysfunction.

AB - Objective: To investigate the regulation of catechol O-methyltransferase (COMT) expression in granulosa cells and assess potential effects of 2-methoxyestradiol (2-ME2) and COMT inhibitors on granulosa cell steroidogenesis and proliferation. Design and Setting: Controlled experimental study in an academic research laboratory. Intervention(s): JC410 porcine and HGL5 human granulosa cell lines were used for in vitro experiments. Effects of 2-ME2 and COMT inhibitor treatment on DNA proliferation and steroidogenesis were assessed by using Hoechst dye and p450SCC-luciferase reporter assays. Effects of dihydrotestosterone (DHT), insulin, and all-trans retinoic acid (ATRA) on COMT messenger RNA expression were investigated by using COMTP1 promoter-luciferase reporter and Northern blot. Main Outcome Measure(s): Granulosa cell steroidogenesis and proliferation following COMP inhibitor and 2-ME2 treatment. Regulation of COMT expression with DHT, insulin, and ATRA. Result(s): 2-Methoxyestradiol had a dual effect on granulosa cell proliferation and p450SCC- luciferase activity; low doses were stimulatory and high doses were inhibitory. Catechol O-methyltransferase inhibitor was associated with up to a 65% increase in JC410 cell number and a maximal 5.6-fold increase in p450SCC-luciferase activity at 20 μmol/L. Dihydrotestosterone, insulin, and ATRA all induced a dose-dependent increase in COMTP1-luciferase transactivation, as well as up-regulated COMT messenger RNA expression in granulosa cells. Conclusion(s): Catechol O-methyltransferase expression in granulosa cells was up-regulated by insulin, DHT, and ATRA. Catechol O-methyltransferase product, 2-ME2, decreased, whereas COMT inhibitor increased granulosa cell proliferation and steroidogenesis. These data suggest that COMT overexpression with subsequent increased level of 2-ME2 may lead to ovulatory dysfunction.

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KW - methoxyestrogen

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