Immunohistochemical detection of estrogen and progesterone receptors in endometriotic tissue

30 formalin-fixed and paraffin embedded and 20 fresh frozen samples of endometriotic tissue were analysed immunohistochemically for the concentration of estrogen and progesterone receptors. In the formalin-fixed and paraffin embedded group only 37% of the samples were estrogen receptor positive whereas 63% were receptor negative. In contrast, we found that 67% of the samples had a positive progesterone receptor status. In the fresh frozen group 60% of endometriotic tissues were estrogen receptor positive and 75% of the tissues had a positive progesterone receptor status. We could not find any correlation between the site or severity of the endometriosis or the hormonal receptor status. We were able to demonstrate that the immunohistochemical detection of hormonal receptors in endometriotic tissues is possible and that better results were obtained if fresh frozen rather than formalin-fixed and paraffin embedded tissues were analyzed.     

Immunohistochemical determination of estrogen and progesterone receptors and DNA flow cytometry in endometrial cancer

Estrogen and progesterone receptor contents (ER, PR) were assessed by an immunohistochemical method and DNA ploidy and S-phase by flow cytometry in frozen endometrial cancer tissue sections from 39 cases. Comparison of the immunohistochemical and cytosol assays showed 81% and 84% concordance in ER and PR contents, respectively. An aneuploid DNA pattern was identified in 30% and a high S-phase fraction was found in 33% of 36 specimens studied. Negative ER status was associated with aneuploid and high S-phase fraction. A similar association was found between PR status and high S-phase fraction. Combined analysis of immunohistochemical receptor status and DNA flow cytometry in the same sample makes it possible to identify two strong predictive factors in endometrial adenocarcinoma.     

子宫肉瘤雌、孕激素受体表达与临床生物学特性及预后的关系

目的　研究子宫肉瘤雌、孕激素受体的表达情况以及其临床生物学意义。方法　选取 1 991年9月～ 2 0 0 1年 9月北京大学人民医院及第三医院收治的子宫肉瘤患者 4 4例 ,选取石蜡病理切片 ,用免疫组化法测定ER、PR表达情况。结果　①ER表达阳性率在子宫内膜间质肉瘤中为 75 % ,子宫平滑肌肉瘤为 31 8% ,子宫恶性苗勒管混合瘤为 2 /6。子宫内膜间质肉瘤ER表达阳性率显著高于子宫平滑肌肉瘤 (P <0 0 5 ) ;②PR表达阳性率在子宫内膜间质肉瘤中为 1 0 0 % ,子宫平滑肌肉瘤为 6 8 2 % ,子宫恶性苗勒管混合瘤为 3/6。子宫内膜间质肉瘤的PR表达阳性率显著高于子宫平滑肌肉瘤及恶性苗勒管混合瘤 (P <0 0 5 ) ;③与临床病理学特征的关系 :ER与PR间呈显著正相关性。无瘤存活者ER、PR表达率显著高于因瘤死亡者 (P <0 0 5 )。核分裂 <1 0个 /1 0HPs的子宫肉瘤PR表达阳性率显著高于核分裂≥ 1 0个 /1 0HPs的子宫肉瘤 (P <0 0 5 )。结论　①子宫内膜间质肉瘤ER、PR阳性率均显著高于子宫平滑肌肉瘤和子宫恶性苗勒管混合瘤 ,子宫内膜间质肉瘤患者可常规进行ER、PR测定 ,根据ER及PR表达情况辅助激素治疗 ;②无瘤存活者ER、PR表达率显著高于因瘤死亡者 ,说明ER、PR表达与子宫肉瘤的预后有关     

RU_(486)对早孕妇女蜕膜组织甾体激素受体及内分泌的影响

目的 了解米非司酮及米索对人早孕蜕膜雌激素受体(ER)、孕激素受体(PR)的表达及血清内分泌水平的影响。方法 取正常早孕、服用米非司酮及服米非司酮配伍米索后各20例的蜕膜,应用单克隆抗体链霉素亲生物蛋白——过氧化酶 (SP) 免疫组织化学方法测定 ER、PR 的表达及血清激素水平的变化。结果米非司酮配伍米索使蜕膜组织中 ER、PR 表达下调,正常早孕组、服米非司酮组及服米非司酮配伍米索组之间差异有显著性。血清雌二醇(E_2)、睾酮(T)、泌乳素(PRL)含量上升,孕酮(P)下降;而对 FSH、hCG 的含量无明显影响。结论 大剂量RU_(486)能竞争地结合蜕膜组织孕激素受体,使早孕期高浓度的内源性孕酮与其受体的正常结合受到干扰,从而达到抗早孕的目的。     

雌激素对子宫内膜癌孕激素受体亚型调控的研究

目的 研究不同剂量雌激素(雌二醇,E_2)对子宫内膜癌细胞系中孕激素受体亚型在蛋白水平的调控,探讨雌激素、孕激素受体亚型与子宫内膜癌的关系。方法 体外培养子宫内膜癌细胞系HEC-IB,分别用含5nm、10nm、20nm雌二醇的培养液分别作用于细胞24、48、72h,以不含雌二醇的培养液培养细胞为自身对照。通过Western blot方法观察不同剂量雌激素对孕激素受体亚型A、B的蛋白水平调控的动态变化,并以乳腺癌细胞系MCF-7为对照。结果 ①HEC-IB细胞分别经5nm、10nm、20nm的E_2作用24、48、72h后,与未加药的细胞相比,不同浓度、时间下E_2对hPRB调控作用均不明显;而对hPRA的作用在E_2 5um时下调,且随时间延长,下调作用越加明显;在10nm的第24、48h下降,其后渐上升,而在E_2 20nm作用72h时基本恢复加药前的水平。②MCF-7细胞分别经E_2 5nm、10nm、20nm作用24、48、72h后,与未加药的细胞相比较,各种浓度及时间下,E_2对hPRA、B有上调作用的趋势;其中10nm的E_2上调作用较为明显,各种浓度的E_2在第48h的上调作用更为显著。结论 E_2对hPR亚型的调控具有细胞特异性、时间及剂量的依赖性。HEC-IB细胞中,各种浓度、时间下E_2对hPRB调控作用均不明显;而对hPRA的调控作用具有下调的趋势。     

Effect of levonorgestrel intrauterine device on human endometrial estrogen and progesterone receptors

Estrogen and progesterone cytoplasmic receptors (ER, PgR) were determined by radiochemical DCC technique in the endometria of 16 normal women at before child bearing age and 6 to 9 months after LNG IUD insertion. The study showed that both ER and PgR were reduced significantly after LNG IUD insertion (P less than 0.01). The decrease of ER and PgR might play an important role in gland reduction and endometrial atrophy. It may be one mechanism of the contraceptive effect and the cause of anemia or spotting between menstruation during the course of LNG IUD insertion.     

Effect of two antiprogestins (mifepristone and onapristone) on endometrial factors of potential importance for implantation

Objective: To investigate the effects of postovulatory administration of antiprogestins on endometrial factors that may be of importance for successful implantation.

Design: Ten women were given 200 mg mifepristone and an additional 10 women 400 mg of onapristone 48 hours after the LH surge in urine (LH+2).

Main Outcome Measure(s): Biopsies were assessed for histologic dating and the immunolocalization of [1] leukemia inhibitory factor,[2] 15-hydroxyprostaglandin dehydrogenase, and [3]the cell proliferation marker Ki 67. Hormonal measurements in blood and urine were used to monitor the effects on the ovarian cycle. Glycodelin (placental protein 14) concentrations were measured in blood taken on LH±12.

Result(s): Treatment with antiprogestins retarded the development of secretory changes without affecting the length of the luteal phase. In addition, there was reduced immunostaining for 15-hydroxyprostaglandin dehydrogenase within glands and a significant reduction in serum levels of glycodelin. Reduced immunostaining for leukemia inhibitory factor also was apparent within glands in biopsies taken on LH+6 of the treatment cycle. Increased Ki 67 immunostaining was observed on both cycle days after treatment, consistent with P antagonism.

Conclusion(s): Administration of mifepristone and onapristone adversely affects uterine receptivity. This adds further evidence to support their potential as a method of postovulatory fertility control.     

雌激素和孕激素对子宫内膜基质细胞血小板反应素-1表达的影响

目的探讨雌二醇(E2)和孕酮(P4)对体外培养的人子宫内膜基质细胞(ES细胞)血小板反应素-1(TSP-1)表达的影响。方法 2007年12月至2008年10月在北京协和医院用不同浓度的E2及P4处理体外培养的ES细胞一定时间(研究组),提取细胞RNA和蛋白,分别用Northern blot和Western blot方法比较研究组和未用激素刺激的对照组之间TSP-1 mRNA和蛋白水平的差异。结果 E2能抑制ES细胞TSP-1的表达,与对照组相比,用10nmol/L的E2处理ES细胞后,其TSP-1 mRNA和蛋白水平分别下降(47.6±6.5)%(P<0.05)和(49.0±8.6)%(P<0.05);高浓度P4(10μmol/L)能诱导ES细胞表达TSP-1,与对照组相比,10μmol/L的P4处理ES细胞后,其TSP-1 mRNA和蛋白水平分别是对照组的(2.1±0.4)倍(P<0.05)和(2.3±0.6)倍(P<0.05);低浓度(10nmol/L)P4能增强E2对TSP-1蛋白表达的抑制作用,但对其mRNA的表达无明显影响。结论雌激素和孕激素能有差别地调节ES细胞TSP-1的表达,这一发现为子宫内...     

子宫内膜癌组织中雌激素及孕激素受体亚型的表达研究

目的探讨子宫内膜癌组织中雌激素受体(ER)亚型mRNA及孕激素受体(PR)亚型mRNA和蛋白表达水平的变化及其意义。方法采用RT-PCR法检测66例子宫内膜癌和30例正常子宫内膜组织ER、PR亚型mRNA的表达,采用蛋白印迹法检测PR亚型蛋白的表达。结果(1)ERαmRNA在子宫内膜癌和正常子宫内膜组织中的表达水平分别是8.00±7.77、3.84±2.57,而ERβmRNA的表达水平分别是4.15±3.55、0.41±0.29,子宫内膜癌组织中ERα、ERβmRNA表达水平均高于正常子宫内膜,两种组织间分别比较,差异均有统计学意义(P<0.05)。(2)PR、PRA和PRB蛋白表达及PR mRNA表达的降低与子宫内膜癌的发生有关(P<0.05),而PRA与PRB蛋白表达的比值和PRB mRNA的表达与子宫内膜癌的发生无明显相关性(P值分别为0.550、0.901)。(3)子宫内膜癌组织中PRB mRNA与ERβmRNA的表达水平呈正相关关系(r=0.43,P<0.01)。结论子宫内膜癌组织中ER亚型mRNA表达上调、PR蛋白和mRNA表达的下调可能参与了子宫内膜癌的发生;PRB mRNA与ERβmRNA表达呈正相关关系。     

Immunohistochemical expression of estrogen and progesterone receptors and outcomes in patients with newly diagnosed uterine leiomyosarcoma

Objective

We assessed the IHC expression of ER and PR and their prognostic significance in uterine leiomyosarcoma (LMS).

Methods

We identified 43 “high-grade” uterine LMS cases from 7/82-7/07 for whom ER/PR IHC analysis was performed at initial diagnosis at our institution.

Results

Disease was confined to the uterine body in 20/43 (47%). Eighteen (42%) of 43 were ER(+); 17/42 (41%) were PR(+). At last follow-up, 33 (77%) had recurred or progressed, and 23 (54%) had died. PR expression was associated with improved progression-free survival (PFS; P = 0.002) and overall survival (OS; P = 0.03) overall; ER expression was not. After adjusting for stage, ER expression was associated with PFS (P = 0.01), not OS (P = 0.3), and PR expression maintained a significant association with PFS (P = 0.002) and approached a significant association with OS (P = 0.05). Neither ER nor PR expression was associated with outcome in cases with disease outside the uterine body. In cases with confined disease, median PFS for ER(+) or PR(+) cases was not reached compared to 16.9 months for ER(−) cases (95% CI: 8.1-25.7; P = 0.03) and 13.5 months for PR(−) cases (95% CI: 5.9-21.1; P = 0.001). Only 1/10 PR(+) cases recurred and died; 9/10 PR(−) cases recurred, and 5 died. Two of 9 ER(+) cases recurred and died; 8/11 ER(−) cases recurred, and 4 died.

Conclusion

ER/PR expression is associated with survival outcomes in patients with high-grade uterine LMS confined to the uterine body. PR expression seems capable of identifying cases confined to the uterine body, which have better outcomes.     

Utility of MIB-1 and estrogen and progesterone receptor in distinguishing between endometrial stromal sarcomas and endometrial stromal nodules, highly cellular leiomyomas

It is difficult to differentiate between an endometrial stromal nodule (ESN) and endometrial stromal sarcoma (ESS) in curettage specimen, and the recommended therapy of endometrial stromal neoplasm is hysterectomy. If we could discriminate ESS from ESN in curettage specimens, there would be an opportunity to treat ESN by local excision rather than by hysterectomy. We analyzed MIB-1 and estrogen and progesterone receptor (ER/PR) expression in a retrospective series of 8 ESSs, 7 ESNs, and 17 highly cellular leiomyomas obtained from hysterectomy specimens. ESSs expressed MIB-1 more frequently than ESNs (P < 0.05), and ESSs had a tendency to express ER less frequently than ESNs (P= 0.08). We observed that in spite of showing MIB-1 expression to some extent, highly cellular leiomyomas usually could not reach ESSs' level and frequency of MIB-1 expression in the current study. Although MIB-1 and ER appear to be promising markers in the differential diagnosis of ESSs, a larger study would be necessary to confirm their validity.     

Effect of levonorgestrel and mifepristone on endometrial receptivity markers in a three-dimensional human endometrial cell culture model

OBJECTIVE: To investigate the effect of levonorgestrel and mifepristone on the expression of endometrial receptivity markers in a three-dimensional endometrial construct. DESIGN: In vitro study. SETTING: University hospital and research laboratory. PATIENT(S): Twelve fertile donors. INTERVENTION(S): Timed endometrial biopsy. MAIN OUTCOME MEASURE(S): Examine the effect of levonorgestrel along with another well-studied fertility-regulating drug, mifepristone, on the expression of endometrial receptivity factors in a three-dimensional stromal and epithelial cell coculture model by immunohistochemistry. RESULT(S): Both epithelial and stromal cells of in vitro endometrial construct showed the presence of estrogen receptor-alpha, estrogen receptor-beta, progesterone receptors-(A+B), vascular endothelial growth factor, leukemia inhibitory factor, interleukin-1 beta, and cyclooxygenase-2, whereas the expression of progesterone receptor-B (AR), integrin alpha(V)beta(3,) and MUC1 were confined to epithelial cells. Mifepristone up-regulated expression of epithelial estrogen receptor-beta and progesterone receptor-B and down-regulated stromal vascular endothelial growth factor and surface molecules MUC1 and integrin alpha(V)beta(3) as observed in vivo. Levonorgestrel had no effect on the expression of endometrial receptivity markers studied. CONCLUSION(S): This in vitro model expresses progesterone-regulated endometrial receptivity factors seen in the physiologic condition. Treatment with levonorgestrel did not affect the expression of these endometrial receptivity markers in contrast to mifepristone. This in vitro model holds the potential to study endometrial receptivity, the embryo-endometrial interaction, and develop new agents for fertility control.     

米非司酮对离体异位与在位子宫内膜雌、孕激素受体含量的影响

目的　探讨子宫内膜异位症(内异症)的异位与在位内膜雌、孕激素受体(ER、PR)含量,及米非司酮对其影响。方法　采用免疫细胞化学法,分析22例内异症患者的在位内膜细胞和其中12例患者的异位内膜细胞体外培养后的ER、PR含量,观察不同浓度米非司酮(1×10-6mol/L和1×10-4mol/L)作用后的变化,并以13例正常子宫内膜作对照。结果　内异症的在位内膜ER、PR含量呈明显周期性变化,分泌早期腺体PR含量显著高于正常子宫内膜［组织化学评分(下同)为2.77±0.32与2.20±0.26,P<0.05］。内异症的异位内膜,增殖期ER(腺体0.65～2.17,间质0.45～1.03)、PR含量(腺体0.55～1.77,间质0.40～1.27)显著低于在位内膜(ER腺体1.50～3.23,间质0.80～1.96;PR腺体1.55～3.34,间质0.98～2.50,P<0.05～0.01);分泌早期无差异;分泌晚期腺体ER含量(3.27±0.31)、PR含量(3.33±0.23)与间质ER含量(1.87±0.31)显著高于在位内膜(分别为0.28±0.11、0.36±0.23和0.26±0.15,P<0.01),而间质PR含量无差异。米非司酮可明显降低内异症的异位和在位内膜ER、PR含量(P<0.01),且米非司酮浓度越高,ER、PR含量降低越明显。结论　内异症的异位和在位内膜ER、PR含量明显不同,米非司酮可下调异位和在位内膜ER、PR的含量。     

The effects of estrogen and progesterone on blood glutamate levels during normal pregnancy in women

Abstract

The purpose of this study was to examine whether changes in estrogen and progesterone levels observed during normal pregnancy influence blood glutamate levels. One-hundred and sixteen pregnant women were divided into three groups based on gestational age: group 1 included women in their first trimester, group 2 included women in their second trimester, and group 3 included women in their third trimester. A single venous blood sample was collected and analyzed for concentrations of estrogen, progesterone, glutamate-pyruvate transaminase (GPT), glutamate-oxaloacetate transaminase (GOT), and glutamate. Concentrations of blood glutamate were significantly lower during the second trimester (p?<?0.001) and third trimester (p?<?0.001). Blood glutamate levels were inversely correlated with levels of estrogen and progesterone throughout pregnancy (p?<?0.001). Levels of GOT and GPT remained stable during the course of pregnancy, apart from a moderate reduction in GPT during the third trimester. Increases in estrogen and progesterone levels during advanced stages of pregnancy were inversely correlated with maternal blood glutamate concentrations. Once a maximal blood glutamate-reducing effect was achieved, any additional estrogen and progesterone had a negligible effect on blood glutamate. This study demonstrates the glutamate-reducing effects of estrogen and progesterone, which is most likely not mediated by a GOT/GPT conversion mechanism.