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Molecular biologic study on the growth inhibition of human endometrial glands.

抑制人子宫内膜腺体生长的分子生物学研究。

基本信息

批准号：
09671670
负责人：
SHIOZAWA Tanri
金额：
$ 2.11万
依托单位：
Shinshu University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671670/
关键词：
endometrium cell cycle cyclin cyclin-dependent kinase (cdk)tumor suppressor gene progesterone p27 p53 apoptosis サイクリンE cdk2

项目摘要

The growth of the glandular cells of the human endometrium is thought to be stimulated by estrogen, and inhibited by progestin. In the present study, to understand the progesterone-induced growth inhibition of the glandular cells of the human endometrium in a view of cell cycle regulatory machineries, following experiments ,were performed. (1) The immunohistochemical expression of cyclins and cyclin-dependent kinases (cdks) of the glandular cells was observed in the proliferative phase, but not in the secretory phase. Among several tumor suppressor gene products, only p27 was immunohistochemically observed in the secretory phase. (2) The expression of cyclins and cdks detected by immunoblotting in cultured endometrial glandular cells was reduced in progesterone treated cells. Conversely, the expression of p27 protein was increased in a dose dependent fashion in those cells treated with progesterone. (3) The expression of p27mRNA examined by Northern blotting analysis did not vary significantly between the proliferative and the secretory phases. (4) To evaluate the effect of progesterone on the proteolysis of p27 in cultured endometrial glands, amount of p27 protein treated with and without progcsterone was evaluated by immunoblotting in every 12 hours (up to 48 hours) after cycloheximide (a protein synthesis inhibitor) was induced. The result showed that the amount of p27 protein was more abundant in those cells treated with progesterone in every time course examined. (5) Forced expression of p27 protein by p27 expression plasmid in cultured endometrial glands induced increased expression of p27 and reduced expression of proliferating cell nuclear antigen (PCNA, an indicator of cell proliferation). In summary, the present study demonstrated that progesterone-induced reduced expression of cyclins/cdks, as well as elevated expression of p27 may be involved in the growth inhibition of endometrial glands.

人子宫内膜腺细胞的生长被认为受到雌激素的刺激，并受到雌激素抑制。在本研究中，为了从细胞周期调节机制的角度了解孕酮诱导的人子宫内膜腺细胞生长抑制，进行了以下实验。(1)细胞周期蛋白和细胞周期蛋白依赖性激酶（cdks）的免疫组化染色在增殖期可见，而在分泌期未见表达。在几种抑癌基因产物中，只有p27在分泌期被化学染色观察到。(2)孕激素处理的子宫内膜腺上皮细胞免疫印迹检测到细胞周期蛋白和cdks的表达减少。相反，在用孕酮处理的那些细胞中，p27蛋白的表达以剂量依赖性方式增加。(3)北方印迹分析显示p27 mRNA的表达在增殖期和分泌期之间无明显差异。(4)为了评估孕酮对培养的子宫内膜腺体中p27蛋白水解的影响，在放线菌酮（蛋白合成抑制剂）诱导后，每12小时（最多48小时）通过免疫印迹评估用和不用孕酮处理的p27蛋白的量。结果表明，在各时间段，孕酮处理的细胞中p27蛋白的表达量均较对照组丰富。(5)在培养的子宫内膜腺体中，通过p27表达质粒强制表达p27蛋白可诱导p27表达增加和增殖细胞核抗原（PCNA，细胞增殖的指标）表达减少。综上所述，本研究表明，炔雌醇诱导的cyclins/cdks表达减少，以及p27表达升高可能参与子宫内膜腺体的生长抑制。