Studies on over-expression of aromatase in leiomyoma of the uterus and its application for conservative management

子宫肌瘤中芳香化酶过度表达的研究及其在保守治疗中的应用

• 批准号: 11671602
• 负责人: SHOZU Makio
• 金额: $ 2.43万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671602/
• 关键词: Aromatase; Leiomyoma of the uterus; In situ estrogen; autocrine mechanism; Molecular biology; プロモーター; 転写制御

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.

在本研究中，我们详细研究了芳香化酶P450在子宫肌瘤中的表达，以确定原位雌激素在子宫肌瘤生长优势中的生理和病理作用。1)我们通过三种不同和互补的方法证明了芳香酶P450在子宫肌瘤组织中的过度表达：用定量竞争RT-PCR方法检测芳香酶P45O转录本的水平；用蛋白质印迹法对子宫肌瘤组织的微体部分进行半定量分析。2)为了鉴定子宫肌瘤组织中表达芳香化酶P450的细胞类型，对组织标本和从新鲜子宫肌瘤组织中释放和培养的肌瘤细胞进行免疫组织化学分析。3)为了确定内源性芳香化酶P450是否在促进子宫肌瘤细胞生长中起作用，我们用WST-1法和胸腺嘧啶核苷掺入法检测了不同浓度的雌激素和雄激素处理的子宫肌瘤细胞W的生长情况。我们证实，子宫肌瘤细胞本身合成雌激素，通过自分泌/内分泌机制促进其生长。4)术前促性腺激素释放激素(GnRH)激动剂治疗可显著抑制子宫肌瘤组织中芳香化酶P450的表达。体外实验表明，促性腺激素释放激素激动剂直接作用于子宫肌瘤细胞，降低芳香酶的表达。GnRH激动剂对原位芳香酶的这种直接抑制作用似乎有助于在GnRH激动剂治疗过程中肌瘤的快速消退。

项目成果

生水真紀夫, 瀬川智也, 上野浩久, 炭谷宏志, 揚惠娟, 井上正樹: "腫瘍とアロマターゼ(1)"産婦人科治療. 78. 452-460 (1999)

Makio Omizu、Tomoya Sekawa、Hirohisa Ueno、Hiroshi Sumiya、Keizan Yang、Masaki Inoue：“肿瘤与芳香酶（1）”妇产科治疗。 78. 452-460（1999）

Murakami K, Nakagawa T, Shozu M, Uchide K, Koike K, Inoue M: "Changes with aging of steroidal levels in the cerebrospinal fluid of women"Maturitas. 33(1). 71-80 (1999)

Murakami K、Nakakawa T、Shozu M、Uchide K、Koike K、Inoue M：“女性脑脊液中类固醇水平随衰老的变化”Maturitas。

Sumitani H,Shozu M,Segawa T,Murakami K,Yang HJ,Simada K,Inoue M: "In situ estrogen synthesized by aromatase P450 in uterine leiomyoma cells promotes cell growth probably via an autocrine /intracrine mechanisnm"Endocrinol.. 141(10). 3852-61 (2000)

Sumitani H，Shozu M，Sekawa T，Murakami K，Yang HJ，Simada K，Inoue M：“子宫肌瘤细胞中芳香酶 P450 合成的原位雌激素可能通过自分泌/内分泌机制促进细胞生长”Endocrinol.. 141（10）

Shozu M, Segawa T, Ueno H, Sumitani H, Yang H-J, Inoue M: "Gynecologic tumors and anomatase (2)"Sanfujinka Chiryo. 78(5)(Japanese). 591-599 (1999)

Shozu M、Sekawa T、Ueno H、Sumitani H、Yang H-J、Inoue M：“妇科肿瘤和异常酶（2）”Sanfujinka Chiryo。

Shozu M,Sumitani H,Murakami k,Segawa T,Yang Hei-Juan,Inoue M.: "Regulation of aromatase activity in bone-derived cells : Possible role of mitogen-activated protein Kinase"J Steroid Biochem Mol Biol. (in press).

Shozu M，Sumitani H，Murakami k，Sekawa T，Yang Hei-Juan，Inoue M.：“骨源细胞中芳香酶活性的调节：丝裂原激活蛋白激酶的可能作用”J Steroid Biochem Mol Biol。