Peter C.M. Young

Cytoplasmic progesterone and estradiol receptors in normal, hyperplastic, and carcinomatous endometria: Therapeutic implications

This study was designed to determine whether the presence of progesterone receptors (PR) and/or estradiol receptors (ER) could be used to predict progestin responsiveness of recurrent or advanced endometrial cancers. We have demonstrated the presence of physicochemically similar cytoplasmic progesterone and estradiol receptors in normal, hyperplastic, and carcinomatous endometria. All normal endometria contained both PR and ER. Seventy-three percent of endometrial hyperplasias were PR(+) and 93% were ER(+). A decreasing concentration of progesterone receptor activity was observed with increasing tumor anaplasia [grade 1, 84% PR(+); grade 2, 55% PR(+); grade 3, 22% PR(+)] and in irradiated tumors. A statistically significant (p less than 0.001) relationship has been demonstrated between the presence of specific cytoplasmic PR and response to progestin therapy in recurrent or advanced endometrial adenocarcinomas. Thus, we conclude that a PR assay may be used to help select the most appropriate therapy for patients with recurrent or advanced endometrial adenocarcinoma.

Steroid receptors and clinical outcome in patients with adenocarcinoma of the endometrium

Progesterone receptor content was measured in tissue samples from 175 patients with endometrial adenocarcinoma by use of the dextran-charcoal method. The estradiol receptor content was determined in 138 of these samples. Ninety-two tumors (52.6%) tested positive for progesterone receptors (greater than 50 fmol/mg cytosol protein) and 111 (80.4%) tested positive for estradiol receptors (greater than 6 fmol/mg). Median follow-up was 27.3 months (range 1 to 152 months). Progesterone receptor status correlated significantly with grade, histology, adnexal spread, age, and recurrence rate in stage I cancer. There was no correlation between progesterone receptor status and clinical stage, myometrial invasion, peritoneal cytology, retroperitoneal lymph node involvement, or spread to the cervix. Estradiol receptor status correlated with adnexal spread and recurrence rate. Recurrence in patients with stage I disease was significantly more common if tumors were negative for progesterone receptor (16 of 43, 37.2%) than if they were positive (four of 57, 7%; p less than 0.001). Recurrence was also more common if tumors were negative for estradiol receptor (seven of 17, 41.2%) than if they were positive (eight of 63, 12.7%; p = 0.02). In recurrent or advanced disease, response to progestin was independent of estradiol receptor content, but tumors positive for progesterone receptors responded significantly more often than those lacking progesterone receptors. Overall survival was superior for patients with progesterone receptor-positive tumors (p = 0.001). Although survival in clinical stages I and II was also superior in patients with lesions positive for progesterone receptors (p = 0.13), there was no statistical difference in survival between patients with progesterone receptor-positive or -negative cancers and surgical stages I and II disease (p = 0.12). Estradiol receptor status had no apparent correlation with survival.

Progesterone binding in human endometrial carcinomas.

Abstract By means of dextran-coated charcoal assay, the capacity of various endometrial cytosol preparations for specific binding of 3 H-progesterone was determined. With the use of an arbitrary value of 50 femtomoles of bound 3 H-progesterone per milligram of cytosol protein as the breaking point between high and low binding capacities, 19 out of 20 normal endometria had high progesterone-binding capacities. Two out of 11 Grade 1, 3 out of 8 Grade II, and 2 out of 4 Grade III endometrial carcinomas showed low binding capacities. All 4 endometrial polyps, 7 out of 9 hyperplastic endometria, and 0 out of 7 nonendometrial gynecologic tumors had high binding capacities. These data suggest a progressive loss of specific progesterone-binding activity from normal endometria to hyperplastic endometria, and from the well-differentiated to the anaplastic forms of endometrial adenocarcinoma. There seemed to be an inverse relationship between age and concentration of progesterone receptors in endometrial adenocarcinomas. All irradiated tumors studied had low progesterone-binding capacity.

Influence of elevated temperature on natural killer cell activity, lymphokine-activated killer cell activity and lectin-dependent cytotoxicity of human umbilical cord blood and adult blood cells

PURPOSE To determine whether hyperthermia is to the benefit or detriment of host immune function, the effect of hyperthermia was evaluated on various functions of T-lymphocytes from human umbilical cord blood and compared to that of adult blood. METHODS AND MATERIALS Nonadherent mononuclear cells from cord blood or adult blood were used as the effector cells. To generate lymphokine activated killer (LAK) cells, effector cells were kept in culture for 5 days in complete medium containing recombinant human interleukin-2. To activate effector cells to become cytotoxic, cells were kept in culture in complete medium containing Con A. Cytotoxicity was determined in a standard 4-h chromium release assay using K-562 human erythroleukemic cells (in the natural killer cell activity assay) or Daudi cells (in the LAK cell activity or Lectin dependent cytotoxicity assay) as targets. For heat effects, cells in complete medium were heated at the desired temperature in a water bath for 1 h. RESULTS Lymphokine-activated killer cell activity, lectin-dependent cytotoxicity and T-cell proliferative capacity were not deficient in human cord blood. Cytotoxic activities of T-cells from adult blood as well as from cord blood can be enhanced at febrile range (< or = 40 degrees C), and were significantly decreased by exposure to 1 h at 42 degrees C. CONCLUSION The febrile responses (< or = 40 degrees C) to infection, in the course of malignant disease and with biological response modifiers treatment, may all be related to host defense mechanisms. Based on these observations, whole body hyperthermia (< or = 40 degrees C), in combination with the appropriate cytokines, may have therapeutic potential in the treatment of neonatal infections and malignancies under certain circumstances. Hyperthermia in febrile range may, therefore, confer an important immunoregulatory advantage to the host. In contrast, tumor killing therapeutic temperature (> 42 degrees C) which inhibits host immunocompetence should probably be used only for local hyperthermia.

Relationship between steroid receptors and response to endocrine therapy and cytotoxic chemotherapy in metastatic breast cancer

The relationship between steroid receptor content and response of metastatic breast cancers to endocrine therapy and cytotoxic chemotherapy has been examined. In 54 advanced breast cancers treated with hormonal therapy, 20 of 29 (69%) ER+, PR+ and 3 of 14 (21%) ER+, PR− tumors responded. Of the nine tumors that lacked both ER and PR, 2 (22%) were hormonally responsive. Based on either the ER or PR content alone, 68% of the PR+ and 54% of the ER+ tumors responded to endocrine therapy. Thus, the addition of PR assay could improve the accuracy of ER assay in selecting a hormonally responsive breast cancer. In 29 advanced breast cancers treated with cytotoxic drugs, the presence of ER and/or PR in the tumors seemed to favor an objective response to cytotoxic chemotherapy. The response rate was about 67% regardless of whether the cancers were ER+ or PR+.

Features associated with survival and disease-free survival in early endometrial cancer

Age, clinical stage, histologic grade, depth of myometrial penetration, adnexal status, peritoneal cytology, and progesterone and estrogen receptor status were available for 139 patients with clinical stage IA, 113, or II endometrial adenocarcioma who had therapy at Indiana University Hospital or St. Vincent Hospital in Indianapolis. These features were analyzed for their association with survival and disease-free survival. Patients treated at Indiana University Hospital were similar to those from St. Vincent Hospital when comparisons were made by X2 test for age, clinical stage, grade, adnexal metastases, peritoneal cytologic results, progesterone receptor status, or estrogen receptor status. However, patients treated at Indiana University Hospital had lesions that were deeper (p = 0.03) than those treated at St. Vincent Hospital. Survival differences were observed for patients with progesterone receptor-rich versus progesterone receptor-poor tumors (p = 0.004), grades 1 and 2 versus grade 3 lesions (p = 0.013), and malignant versus benign peritoneal cytologic results (p = 0.01). Differences in disease-free survival were observed for those patients with adnexal metastases versus those with no adnexal disease (p = 0.002), those with estrogen receptor-rich versus estrogen receptor-poor tumors, outer third myometrial invasion (p = 0.002), and patients with clinical stage I versus clinical stage II disease (p = 0.03). A stepwise Cox proportional hazards model was constructed to determine correlates of disease-free survival. In the final model, grade (p = 0.0002), peritoneal cytologic results (p = 0.0002), progesterone receptor status (p = 0.004), and age as a continuous variable (p = 0.008) were most closely associated with disease-free survival.

Estrogenic effects of the antiprogestin onapristone (ZK98.299) in the rodent uterus

OBJECTIVE Our purpose was to assess the estrogenic action of onapristone (ZK299). STUDY DESIGN Three rodent models of estrogen action in the uterus were used. Deoxyribonucleic acid synthesis in the uterine epithelium of neonatal mice was determined by thymidine autoradiography. In adult ovariectomized mice uterine wet weight and progesterone receptor and estrogen receptor concentrations were determined. In immature rats uterine deoxyribonucleic acid synthetic activity was determined by thymidine autoradiography, epithelial hypertrophy and stromal edema were assessed histomorphometrically, and complement C3 protein synthesis was assessed by metabolic labeling in vitro. The effects of ZK299 were challenged with the antiestrogens ICI164,384 and tamoxifen. The ability of ZK299 to displace tritiated estradiol from the estrogen receptor was assessed in cytosolic preparations from mouse uterus. RESULTS In the neonatal mouse ZK299 stimulated epithelial deoxyribonucleic acid synthesis; two other antiprogestins, RU486 and ZK98.734, had no effect. Three daily injections of ZK299 at 10 micrograms/gm body weight to 6-week-old ovariectomized mice increased uterine progesterone receptor 42%; this effect was blocked by ICI164,384. In another experiment three daily doses of ZK299 (20 micrograms/gm) to 10-week-old ovariectomized mice increased progesterone receptor concentration by 63% and uterine wet weight by 42%. In 21-day-old rats a single injection of ZK299 increased uterine epithelial deoxyribonucleic acid synthesis; this effect was blocked by tamoxifen. Both ZK299 and tamoxifen increased epithelial cell height and thymidine labeling in the stroma. ZK98.734 had no effect on epithelium or stroma. ZK299 also stimulated synthesis of complement C3 by uteri of immature rats. In competitive binding assays ZK299 exhibited weak relative binding affinity (0.05%) for mouse uterine estrogen receptor. CONCLUSIONS ZK299 can act as a weak estrogen in the rodent uterus, most likely through a direct, low-affinity interaction with the estrogen receptor. Because estrogens may increase the risk for endometrial, breast, and liver cancer, caution is warranted in long-term administration of this drug to women.

Effect Of Metal Ions On The Binding Of 17β-Estradiol To Humai Endometrial Cytosol

The influence of zinc and other metal ions on the binding of 3 H-17β-estradiol to human endometrial cytosol was studied. Zn 2+ began to interfere with estrogen binding when its concentration in the cytosol exceeded 50 μM. At a concentration of 5 mM, all of the specific binding of 17β-estradiol, including more than 50% of the nonspecific binding of the hormone, was destroyed. Analysis of the binding data revealed that one possible site of action of the cations on the binding protein might be the sulfhydryl group(s) of the 17β-estradiol binding site. The inhibition of 17β-estradiol binding brought about by Zn 2+ was partially abolished by dithiothreitol. Among the metal ions tested, Cu 2+ was found to be the most potent inhibitor, followed by Cd 2+ , Zn 2+ , and Pb 2+ . At 1 mM, Mn 2+ , Ba 2+ , Ca 2+ , and Mg 2+ had little effect, but at 5 mM, their inhibitory action became more appreciable. K + and Na + had no effect on 17β-estradiol binding. The stimulatory effect of 5 mM Zn 2+ , Ca 2+ , Mg 2+ , and K + on the binding of 3 H-l7β-estradiol to a macromolecular fraction from bovine endometrium was not observed in human endometrial cytosol.

Progesterone and dexamethasone inhibition of uterine epithelial cell proliferation : studies with antiprogesterone compounds in the neonatal mouse

Progestins and glucocorticoids inhibit uterine epithelial cell proliferation. Earlier studies showed that in the neonatal mouse, dexamethasone (Dex) was at least 100-fold more potent than progesterone (P) as an inhibitor of epithelial DNA synthesis. Using steroid autoradiography, we now show that the uterine cells of the neonatal mouse have receptors for both progestins (PR) and glucocorticoids (GR). Since it is known that P can interact with the GR it is possible that even a weak interaction might mediate the inhibitory effects of large doses of P. Therefore, we examined the receptor pathway specificity of the P response in neonatal mice using antiP steroids with reportedly strong antiglucocorticoid activity, RU486, or weak antiglucocorticoid activity, ZK98.734 (ZK734). Both RU486 and ZK734 exhibited strong binding activity in a PR assay performed on cytosolic preparations from adult mouse uteri. For murine thymic GR, the relative binding activity of RU486 was 12-fold that of ZK734. The high PR binding activity was reflected by the in vivo dose-response of the antagonists administered in conjunction with P; either antagonist completely blocked the inhibitory effect of P on uterine epithelial DNA synthesis when it was administered at one-tenth the dose of P. On the other hand, blockade of the inhibitory effect of Dex only occurred when the dose of antagonist was 10-fold the dosage of Dex. There were no significant differences between the two antagonists in blocking the effects of either P or Dex. These results indicate that progestins and glucocorticoids act through their own receptor systems to inhibit DNA synthesis in the uterine epithelium of the neonatal mouse. However, because of the considerable antiglucocorticoid activity of ZK734, a potential role for the interaction between P and the GR cannot be ruled out by these experiments.

Serum unconjugated estriol in normal and abnormal pregnancy

A radioimmunoassay with the use of less than 0.5 ml. of pregnancy serum has been employed to determine the levels of unconjugated estriol in the second half of pregnancy. Sephadex LH-20 was used to separate estriol from estradiol and estrone. In the assay, the bound hormone was separated from the unbound hormone by dextran-coated charcoal. The water blank was below the sensitivity of the assay. Serum unconjugated estriol values revealed a rise in the mean estriol from 4.2 ng- per milliliter at 20 weeks to 14.9 ng. per milliliter at term. In pregnancies complicated by anencephaly and fetal death, the levels were significantly below the normal. In 5 mildly affected diabetic patients, the serum estriol values remained normal and there were no perinatal deaths. The outcome in 8 complicated by hypertension, anemia, leukemia, the nephrotic syndrome, and rheumatic heart disease is described.