| Project/Area Number |
11671602
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Kanazawa University |
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Principal Investigator |
SHOZU Makio University Hospital, Kanazawa University, Assistante Professor, 医学部・附属病院, 助教授 (30226302)
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| Co-Investigator(Kenkyū-buntansha) |
SEGAWA Tomoya University Hospital, Kanazawa University, Assistant Professor, 医学部・附属病院, 助手 (40301197)
KOIKE Kouji Obstetrics and Gynecology, Kanazawa University, Associate Professor, 医学系研究科, 助教授 (70225340)
INOUE Masaki Obstetrics and Gynecology, Kanazawa University, Professor, 医学系研究科, 教授 (10127186)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Aromatase / Leiomyoma of the uterus / In situ estrogen / autocrine mechanism / Molecular biology / プロモーター / 転写制御 |
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| Research Abstract |
In the this study we characterized in detail the expression of aromatase P450 in leiomyomas to determine the physiological and pathological role of in situ estrogen in the growth advantage of leiomyomas.
1) We demonstrated over-expression of aromatase P450 in leiomyoma tissues in comparison to the surrounding myometrium by three different and complementary methods: The levels of aromatase P45O transcripts were quantitated by quantitative competitive RT-PCR The amount of protein was semi-quantitated by Western blot analysis using microsomal fractions of leiomyoma tissues. Enzyme activity was measured by tritiated water-releasing assay using microsomal fractions.
2) To identify a cell type that express aromatase P450 in leiomyoma tissues, immunohistochemical analysis was performed on the tissue samples and the leiomyoma cells that were released and cultured from the fresh leiomyoma tissues. Strong immunoreactivity was localized in the cytoplasm of leiomyoma cells.
3) To determine whether endogenous aromatase P450 plays a role in the growth promotion of leiomyoma cells, we evaluated the cell W growth of leiomyoma cells treated with various concentration o of estrogens as well as androgens using a WST-1 assay and thymidine incorporation assay. We confirmed mat leiomyoma cells per se synthesize estrogen, which promotes their growth via an autocrine/intracrine mechanism.
4) The expression of aromatase P450 in leiomyoma tissues was profoundly suppressed by preoperative gonadotrophin releasing hormone (GnRH) agonist therapy. In vitro experiment indicated that GnRH agonist acted directly on the leiomyoma cells to reduce the expression of aromatase. This direct inhibitory action of GnRH agonist on in situ aromatase seems to contributes to the rapid regression of leiomyoma during GnRH agonist therapy.
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