Action Mechanisms and Responsiveness for Steroid Hormones in Human normal Endometrium and Endometrial Cancer.

（1）实验模型的材料：20个正常内皮素的案例，从用紫外线纤维的操作材料中提取出来，是实验的主要文化。这些材料是有用的，可用于观察体外的激素行为，但对于免疫化学的测定是不成功的。九个细胞线，已被证实来自一个内皮癌的案例，在这项研究中使用过。（2）受体分析：从正常饮食周期中提取正常内皮素的七种情况，从操作材料中提取31种内皮素的癌症方法，被用于检测雌激素和催乳素受体的免疫化学方法，并使用单克隆抗体来对抗雌激素和催乳素受体。此外，还通过生物化学和免疫化学程序检测到了9个内皮癌细胞的受体含量和本地化。通过免疫化学学保持变化的正常内计量检测的受体含量 ... More d =雌激素和孕激素在月经周期期间的状态。The incidence of receptor positive cases in endometrial cancer tissues were decreased corresponding to the loss of histological differentiation。The positive rates of both estrogen and progesterone receptors of G1， G2 and G3 endometrial adenocarcinoma tissues were 8/11 （73%）， 3/13 （23%） and 0/4 （0%）， respectively。Ishikawa细胞是从一个良好的不同的内皮瘤可能的雌激素和催乳素受体中衍生出来的，它们显示出对雌激素和程序的响应，并通过改变参数的不同。在核免疫化学物质中发现了雌激素和催乳素受体的局部化，并确认Gorski和Greene的一步理论的正确性，从而影响了类固醇激素的作用机制。Ishikawa细胞与3 H-R5020的全细胞绑定试验对Ishikawa细胞的绑定进行了测量，该试验被添加到作为剂量和时间依赖性指示器的雌二醇。理想的estradiol范围为10 nM。（3）雌二醇和程序操作的各种参数。（1） Cell Proliferation and Cycle ：Ishikawa Cell Proliferation of Ishikawa Cell s在低血清条件下的雌二醇添加物被刺激。S相细胞对抗支架检测的比例-BrDU单克隆抗体免疫化学上也很清楚地包含在10 nM的雌二醇中。（2） 17 beta雌二醇脱氢酶（E2 DH）活性： The activity of E2 DH as a Parameter of progestin action of normal endometrium which contains the functional progesterone receptor were found in Inshikawa endometrial cells. Ishikawa细胞的这些活动与正常内皮素比较，但通过向文化媒体添加的雌二醇和程序的同时刺激。（3）糖原代谢：正常内皮素作用的这些参数也在内皮素癌症细胞中被调查。糖原合成酶（GS）的活动和石川细胞的糖原含量被程序的添加剂刺激（P4， MPA， R5020和Org 2058）。RU 486的这些对程序的影响完全被废除，其中一个对程序员受体的竞争性试剂。Progestins和RU 486可能需要考虑通过葡萄糖皮质激素受体而不是通过程序一个受体来行动。两种葡萄糖皮质激素（Cortisol、Dexamethasone），也被添加到文化媒介对糖元代谢没有影响，确认Ishkawa细胞的程序员受体是功能性的。在对比度下，石川细胞的糖原磷酸酶的活动已被过程降解。（4） Autocrine and paracrine actions of local growth factor EGF（Epidermal growth factor） is well known as local growth factor which may be able to stimulate the proliferation of a various types of cells。据报道，正常内皮素（EGFR）的受体因雌二醇和催乳素状态而发生了变化，建议成为类固醇激素的参数。在这项研究中，用免疫细胞化学和生物化学方法检测到的内皮细胞EGFR。EGFR的局部化，由抗EGFR单克隆抗体在细胞膜上发现，并在EGF添加到培养媒体后直接在内部化。Ishikawa细胞和HEC-50细胞的EGF绑定数量，根据125个I-EGF与未标记的EGF的竞争性分析进行了评估，分别为2、500和11 000个站点/细胞，准确地衡量。Ishikawa细胞的产生是由EGF的附加物模拟的，以培养媒体在无血清条件下的剂量依赖性标记。Less（低）

(1) Materials for experimental model : Twenty cases of normal endometrium, had been taken from operation material with uterine fibromas, were primarily cultured for the experiments. These materials were useful for observations of hormonal actions in vitro, but were not successful for immunocytochemical assay. Nine cell lines, had been established from the cases of endometrial cancer, were used in this research.(2) Receptor assay : Seventy cases of normal endometrium taken from the patient with normal menstrual cycle, and 31 endometrial cancer tissues from operation materials, were stained with immunocytochemical methods for the detection of estrogen and progesterone receptors, using the monoclonal antibodies against to estrogen and progesterone receptors. The receptor contents and localization of 9 endometrial cancer cells also detected by both biochemical and immunocytochemical procedures. The receptor contents of normal endometrium detecting by immunocytochemical staining were change … More d by estrogen and progesterone status during menstrual cycle. The incidence of receptor positive cases in endometrial cancer tissues were decreased corresponding to the loss of histological differentiation. The positive rates of both estrogen and progesterone receptors of G1, G2 and G3 endometrial adenocarcinoma tissues were 8/11 (73%), 3/13 (23%) and 0/4 (0%), respectively. The Ishikawa cells, derived from a well differentiated endometrial adenocarcinoma possess estrogen and progesterone receptors are showen to respond to estrogens and progestins by changing a variety of parameters. The localization of estrogen and progesterone receptors are found in the nuclei by immunocytochemical stainings, confirming the correctness of Gorski and Greene's one step theory involving the action mechanisms of steroid hormones. The amounts progesterone binding of Ishikawa cells measured by whole cell binding assay for 3H-R5020 were increased by the addition of estradiol to the culture media as dose and time dependent manner. Optimal concentration of estradiol were 10nM.(3) Various parameters for estradiol and progestin action.(1) Cell proliferation and cycle : The cell proliferation of Ishikawa cells were stimulated by the addition of estradiol at 10nM under low serum condition. The proportionality of the S-phase cells detection by the stainings for antiーBrdU monoclonal antobodiesimmunocytochemically also clearly increased by the estradiol at 10nM.(2) 17 beta estradiol dehydrogenase (E2DH) activity : The activity of E2DH as a parameter of progestin action of normal endometrium which contains the functional progesterone receptor were found in Inshikawa endometrial cancer cells. These activity of Ishikawa cells were lower compared with normal endometrium but were stimulated by the simultaneous addition of estradiol and progestin to the culture media.(3) Glycogen metabolism : These parameter of progestin action of normal endometrium were also investigated in endometrial cancer cells. The activity of glycogen synthetase (GS) and glycogen content of Ishikawa cells were stimulated by the additon of progestins (P4, MPA, R5020 and Org 2058). These effects of progestins were completely abolished by RU486 which is competitive reagent on progesterone receptor. Progestins and RU486 may be thought to act via not progesterone receptor but glucocorticoid receptor. Two glucocorticoids (Cortisol, Dexamethasone), had been also added to the culture media d no effect on glycogen metabolism, confirming the progesterone receptors of Ishkawa cells were functional. On the contrast, the activity of glycogen phosphorylase of Ishikawa cells were decreased by progestins.(4) Autocrine and paracrine actions of local growth factorsEGF (Epidermal growth factor) is well known as local growth factor which may be able to stimulate the proliferation of a various types of cells. It has been reported that the receptors of EGF (EGFR) of normal endometrium are changed by estradiol and progesterone status, suggesting to be parameters of steroid hormones. In this study, EGFR of endometrial cancer cells were detected by immunocytoctemical and biochemical methods. The localization of EGFR, stained by anti-EGFR monoclonal antibodies were found on cell membrane and internalized immediately after the addition of EGF to culture media. The amounts of EGF bindings of Ishikawa cells and HEC-50 cells, measured by competitive assay of 125I-EGF with unlabeled EGF, were 2,500 and 11,000 sites/cell, respectively. The proliferations of Ishikawa cells were simulated by the addition of EGF to culture media at dose dependent manner under serum-free conditions. Less