eNOS expression is associated with the estrogen and progesterone receptor status in invasive breast carcinoma

In this study we investigated the immunohistochemical expression of eNOS and nNOS in 80 invasive breast carcinomas. Since NO is known to influence apoptosis and angiogenesis we also determined the apoptotic index of the tumor cells by the TUNEL method and tumor angiogenesis by immunostaining the sections with FVIII-related antigen and assessing the number of positively stained vessels. Respectively, 65% and 11% of the cases expressed cytoplasmic positivity for eNOS and nNOS. The mean apoptotic index of the tumors was 0.60%. eNOS or nNOS expression did not associate with the apoptotic index (p=0.36 and p=0.58, respectively). Cases with a positive estrogen or progesterone receptor status were significantly more often eNOS positive than receptor negative cases (p=0.012 and p=0.015, respectively). No association was found between the estrogen or progesterone receptor status and nNOS expression. Neither eNOS nor nNOS expression was associated with vascular density, tumor grade or the TNM status of the tumors. The results show that eNOS and nNOS are expressed in breast carcinomas possibly contributing to their NO synthesis. No association with apoptosis or angiogenesis was detected. eNOS expression was, however, associated with the positive estrogen and progesterone receptors status suggesting that its synthesis might be regulated by hormonal stimulation.     

Variation of estrogen and progesterone receptor status in breast cancer after tamoxifen therapy

In the present paper we have studied the quantitative variations in estrogen (ER) and progesterone receptor (PR) content of breast cancer induced by tamoxifen. In addition to receptors, hormonal levels of estradiol, progesterone, prolactin, FSH, LH and testosterone were also measured. The cases included in our study were consecutively selected among those breast cancers in which an aliquot of the tissue sample sent for analysis of the steroid receptors was positive for cancer and also found to have at least one of the steroid receptors positive, not only in the biopsy but also in the surgical specimen. Following this criterion, we finally collected 14 cases of breast cancer treated daily with 30 mg of tamoxifen during an interval of 3 weeks from the initial biopsy to the final surgery. From our results we can conclude that tamoxifen reduced significantly the ER concentration while no changes were observed in PR values. Concerning hormones, while in premenopausal patients tamoxifen induced a rise in plasma estradiol, in postmenopausal women the only modification observed was a decrease in plasma FSH. The variation in steroid receptor content under tamoxifen therapy may also contribute to the evaluation of the hormone dependency of gynecologic malignancies.     

乳腺癌肿瘤组织不同部位雌孕激素受体的表达

目的 探讨乳腺浸润性导管癌肿瘤组织中不同部位雌、孕激素受体表达情况.方法 收集32例手术切除乳腺浸润性导管癌肿瘤标本,于每例肿瘤标本4个不同部位取材,用免疫组化方法检测各部位雌、孕激素受体表达情况.结果 肿瘤组织不同部位雌、孕激素受体检测结果一致性好,最好Kappa值分别为0.789和0.810,最差Kappa值分别为...     

Short-term biologic response to withdrawal of hormone replacement therapy in patients with invasive breast carcinoma

BACKGROUND: The biologic effect of continuing hormone replacement therapy (HRT) after a diagnosis of breast carcinoma is unclear. The goal of rhe current study was to determine the short-term effect of HRT withdrawal on invasive breast carcinoma using biologic surrogate markers of tumor response. METHODS: The study was performed between 1996 and 2000 and comprised 140 women who had been using HRT at the time of breast carcinoma diagnosis by core needle biopsy. The breast tumors were removed a median of 17 days later (range, 2-31 days). Of these women, 125 women stopped HRT at the time of core needle biopsy and 15 continued to receive HRT until surgery. In addition, 55 women with breast carcinoma from the same time period, who were not receiving HRT at diagnosis, were studied. Changes in expression of Ki-67 (a measure of epithelial cell proliferation), progesterone receptor (PR), p27KIP-1 (a cyclin-dependent kinase inhibitor), and cyclin D1 (a cell cycle-related protein) were determined by immunohistochemistry on paired sections of the core needle biopsy and surgical specimens from each patient. RESULTS: In women who stopped HRT, a significant decrease in Ki-67 expression was observed between core needle biopsy and surgery in estrogen receptor (ER)-positive (n = 106; P < 0.001), but not in ER-negative tumors (n = 19; P = 0.58), with an associated reduction in PR (P < 0.001) and cyclin D1 expression (P < 0.001) and an increase in p27KIP-1 (P = 0.03). These changes in Ki-67 and PR expression occurred irrespective of c-erb-B2 status. No change was observed in any parameter in the other groups of patients. CONCLUSIONS: ER-positive invasive breast carcinomas demonstrated a favorable biologic response to withdrawal of HRT. Therefore, HRT should be stopped at the time of diagnosis and was subsequently contraindicated.     

乳腺浸润性导管癌及良性病变中ER基因的5''''CpG岛甲基化状态

目的：探讨ER基因5＇CpG岛甲基化在原发性乳腺癌组织中与ER蛋白失表达的关系及其在肿瘤进展中的作用。方法：用限制性酶、PCR检测45例乳腺浸润性导管癌及15例乳腺良性病变雌激素受体（estrogen receptor，ER）基因中5’CpG岛甲基化状态，甲基化检测结果与免疫组织化学方法检测的ER蛋白表达结果进行比较。结果：在45例乳腺浸润性导管癌中，有20例发生甲基化（44％），其中4例在2个酶切位点均发生甲基化，而其他的只在HpaⅡ位点发生甲基化。15例乳腺良性病变中，无1例发生甲基化。82％ER阳性的乳腺癌组织显示在ER基因中HpaⅡ、HhaⅠ位点均未发生甲基化，而61％ER阴性的乳腺癌组织显示在ER基因中HpaⅡ和（或）HhaⅠ位点发生甲基化，提示免疫组化ER蛋白表达与ER基因5’CpG呈负相关（P〈0．05）。结论：人类乳腺浸润性导管癌中ER基因的5’CpG岛的甲基化与ER蛋白的表达呈负相关，且随组织学分级升高，甲基化发生率增加。     

Tumor grade and progesterone receptor status predict 21-gene recurrence score in early stage invasive breast carcinoma

Purpose

Agents targeting programmed death receptor 1 (PD-1) or its ligand (PD-L1) have shown antitumor activity in the treatment of metastatic breast cancer (MBC). The aim of this study was to assess the activity of avelumab, a PD-L1 inhibitor, in patients with MBC.

Methods

In a phase 1 trial (JAVELIN Solid Tumor; NCT01772004), patients with MBC refractory to or progressing after standard-of-care therapy received avelumab intravenously 10 mg/kg every 2 weeks. Tumors were assessed every 6 weeks by RECIST v1.1. Adverse events (AEs) were graded by NCI-CTCAE v4.0. Membrane PD-L1 expression was assessed by immunohistochemistry (Dako PD-L1 IHC 73-10 pharmDx).

Results

A total of 168 patients with MBC, including 58 patients with triple-negative breast cancer (TNBC), were treated with avelumab for 2–50 weeks and followed for 6–15 months. Patients were heavily pretreated with a median of three prior therapies for metastatic or locally advanced disease. Grade ≥ 3 treatment-related AEs occurred in 13.7% of patients, including two treatment-related deaths. The confirmed objective response rate (ORR) was 3.0% overall (one complete response and four partial responses) and 5.2% in patients with TNBC. A trend toward a higher ORR was seen in patients with PD-L1+ versus PD-L1? tumor-associated immune cells in the overall population (16.7% vs. 1.6%) and in the TNBC subgroup (22.2% vs. 2.6%).

Conclusion

Avelumab showed an acceptable safety profile and clinical activity in a subset of patients with MBC. PD-L1 expression in tumor-associated immune cells may be associated with a higher probability of clinical response to avelumab in MBC.

     

Epithelial proliferation and hormone receptor status in the normal post-menopausal breast and the effects of hormone replacement therapy.

The proliferation rate (as assessed by Ki67 expression) and expression of oestrogen-regulated progesterone receptor (PR) was studied in normal post-menopausal breast epithelium. Normal breast epithelium from patients receiving hormone replacement therapy (HRT) at the time of surgery containing either oestrogen alone (E2) or oestrogen and progesterone combined activities (E2 + P) was also studied, as HRT has been linked to an increased breast cancer risk. Samples of breast tissue, containing normal epithelium, from 185 patients undergoing surgery for benign or malignant disease were immunocytochemically stained for PR and Ki67. The percentage of labelled cells was expressed as the labelling index (LI). The median Ki67 LI in normal post-menopausal breast epithelium was 0.19 and median PR LI was 4.75, and both were unaffected by patient age, duration of menopause or if the tissue sample originated from a breast with benign or malignant disease. Proliferation did not alter significantly in patients taking HRT (P = 0.61); however, PR expression was up-regulated in both E2 and E2 + P users (P = 0.01). The dose and duration of HRT had no effect on either parameter. A possible attenuation of sensitivity to oestradiol-induced proliferation but not to PR expression occurs in the post-menopausal breast.     

Pattern of metastases in human breast carcinoma in relation to estrogen receptor status

The distribution of metastases at the first recurrence of breast cancer was studied in 57 estrogen receptor (ER) positive and in 23 ER negative patients, who constituted a subset of 460 patients with operable breast cancer. The pattern of metastases with respect to localization of metastases and the dominant site of first recurrence was similar in patients with ER positive and ER negative tumours. The recurrence-free survival (RFS) and the overall survival were associated with the ER status in the 460 patients. ER positive patients had both a significantly longer RFS (p = 0.0024) and survival (p = 0.0001) compared to ER negative patients. Survival after recurrence was prolonged in patients with soft tissue recurrences only, and the proportion of dead patients was highest in receptor negative patients with metastases to bone and viscera. In conclusion, we could not demonstrate that ER positive and negative tumours have a propensity for recurrence at specific sites.     

Tumour steroid hormone synthesis and estrogen receptor status in breast cancer patients

Summary The capacity of breast cancer to synthesise active androgens and estrogens has been related to estrogen receptor (ER) status in 79 postmenopausal patients with breast cancer. Although there was no quantitative relationship between levels of ER and steroid metabolism in ER positive tumours, there was (a) a positive correlation between estrogen synthesis and ER positivity and (b) increased androgen synthesis and ER negativity. This may imply an inherent difference in the handling of hormones in ER positive and negative tumours. Address for reprints: R.C. Mason, University Department of Clinical Surgery, Royal Infirmary, Edinburgh EH39YW, United Kingdom.     

Solving the dilemma of the immunohistochemical and other methods used for scoring estrogen receptor and progesterone receptor in patients with invasive breast carcinoma

BACKGROUND: The authors attempted to resolve the dilemma posed by the lack of unanimity concerning the optimal immunohistochemical (IHC) method for determining and scoring estrogen receptor (ER) and progesterone receptor (PR). METHODS: Sections for IHC were prepared from paraffin embedded tumor samples from 402 patients with lymph node positive breast carcinoma who had biochemical receptor values (obtained with the dextran-coated charcoal [DCC] method) and who were enrolled in a prospective, randomized trial (National Surgical Adjuvant Breast and Bowel Project protocol B-09). IHC receptors were scored independently by two observers according to percent, intensity, and any-or-none algorithms. Results from these evaluations and from two computer-assisted evaluations, DCC, and common pathologic characteristics were analyzed for optimum splits for positive reactions in univariate and multivariate analyses using a tree-structured model. Concordance, sensitivity, and specificity were determined between the DCC method and all other methods. RESULTS: Interobserver agreement and concordance between the DCC method and the other methods and among the methods were high. Univariate analyses revealed that a positive ER score obtained with all methods was related significantly to overall survival (OS) at 5 years and at 10 years. Results related to PR scores and disease-free survival and recurrence-free survival were less consistent. In multivariate analysis, it also was found that all methods for scoring ER predicted a better prognosis for OS in patients with an unfavorable lymph node status at 5 years and 10 years. Patients in a favorable lymph node status group were discriminated further by nuclear grade. CONCLUSIONS: All IHC methods for scoring ER appeared valid as prognostic indicators of OS in patients with positive lymph nodes. The any-or-none IHC method, by virtue of its simplicity, represents an appropriate choice for practical use.     

Prevalence of androgen receptors in invasive breast carcinoma and its relation with estrogen receptor,progesterone receptor and Her2/neu expression

Background and aimsAlthough Breast carcinoma had many targeted biomarkers for its treatment, however, it is a heterogeneous disease with different outcomes and need new markers especially for the triple negative group when estrogen receptor, progesterone receptors and Her2/neu are negative. Androgen receptor is a new target with unclear role. The aim of this study was to examine the prevalence of androgen receptors in invasive breast cancer and tries to elucidate its relation to some well recognized clinicopathological and immunohistochemical markers.Materials and methodsOne hundred and fifty cases of invasive breast carcinoma were evaluated for type, grade and stage and studied immunohistochemically for estrogen receptor, progesterone receptor, Her2/neu and androgen expression. Androgen receptor expression was correlated with histopathological factors and the three studied markers separately then the studied cases were classified into three groups according to estrogen, progesterone receptor and Her2/neu expression and correlated with androgen receptor expression.ResultsAndrogen receptor was expressed in 71% of breast cancer cases. Its expression is associated significantly with both the stage and the grade. Also it was significantly associated with estrogen receptor and Her2/neu expression. It was expressed in a significant number of triple negative breast carcinoma, in Her2/neu positive cases and in estrogen negative cases which indicate that androgen receptor could be a new target for the treatment of these groups.ConclusionsAlthough the impact of androgen receptor on breast cancer outcomes had not been clearly established, this result may provide evidence that androgen receptor is a good prognostic and predictive marker.     

Dietary intake of isoflavones and breast cancer risk by estrogen and progesterone receptor status

Epidemiological and experimental studies suggest that isoflavones may protect against breast cancer by acting as estrogen agonists or antagonists. A case-control study was conducted in southeast China in 2004–2005 to examine the association between dietary isoflavone intake and breast cancer risk by estrogen receptor (ER) and progesterone receptor (PR) status. The incident cases were 756 female patients with histologically confirmed breast cancer. The 1,009 age-matched controls were healthy women randomly recruited from outpatient breast clinics. We assessed isoflavone intake by face-to-face interview using a validated and reliable food-frequency questionnaire and obtained tumor ER and PR status from pathologic reports. Compared with women in the lowest intake quartiles, those in the highest quartile of total isoflavone intake had a reduced risk of all receptor status subtypes of breast cancer with a dose-response relationship. The adjusted ORs (95% CIs) were 0.39 (0.27–0.58) for ER+, 0.32 (0.21–0.49) for ER−, 0.43 (0.29–0.64) for PR+, and 0.30 (0.19–0.45) for PR− (P for trend <0.001). These inverse associations existed in both pre- and post-menopausal women after stratification. Stronger evidence of a protective effect of high isoflavone intake was observed for breast cancer tumors with concordant rather than discordant receptor status; i.e., those with ER+/PR+ (OR 0.39, 0.26–0.59) and ER−/PR− (OR 0.28, 0.17–0.44). The finding that isoflavones protect against all tumor subtypes of breast cancer have biological plausibility, being supported by evidence from experimental studies. Future studies are required to fully understand the complex regulation of isoflavone on breast cancer by tumor hormone status.     

Oestrogen and progesterone receptor status of individual foci in multifocal invasive ductal breast cancer

Widespread acceptance of breast conserving surgery for early breast cancer has led to renewed interest in multifocality, which is seen in 13-63% of breast cancers. According to current guidelines, oestrogen/progesterone receptor status is assessed on the sample obtained at initial core biopsy or the main tumour focus in multifocal breast cancer (more than one distinct tumour focus in a quadrant). We assessed receptor status of individual foci in multifocal breast cancer. Mastectomy specimens for 18 cases of multifocal breast cancer were identified. Immunohistochemical staining for oestrogen and progesterone receptors was performed on all tumour foci. On histological examination 11 patients demonstrated two independent tumour foci, three demonstrated three foci and four demonstrated four foci. Minor differences in oestrogen receptor score were seen between foci (attributed to the subjective nature of the scoring system), which did not affect the overall positive/negative classification. Sixteen patients (88%) were oestrogen receptor-positive. Progesterone receptor staining showed more variability between foci in two patients but, since the tumours were oestrogen receptor-positive this would not have affected clinical decision-making. No major differences in oestrogen receptor status between multiple tumour foci in the same quadrant were found in this pilot study.     

Comparison of estrogen and progesterone receptor status to lymphocyte immunity against tumor antigens in breast cancer patients

Summary Estrogen (ER) and progesterone receptor (PgR) content of tumors were determined by both the dextrancoated charcoal (DCC) cytosol and immunocytochemical assays (ICA), and these hormone receptor results were compared to lymphocyte immunity against tumor antigen(s) for 52 breast carcinoma patients. Hormone receptor analysis by both methods demonstrated that 60% of the patients' tumors had ERs, while 44% were positive for PgRs. The ICA procedure was more sensitive than the cytosol technique for determining PgR content of the tumors. This increased sensitivity was not observed for ER by ICA. Patient age, tumor size, and nodal status were not related to the ER and PgR receptor status. A total of 21/52 (40%) of the patients had positive lymphocyte immunity against tumor antigen. This immunity was independent of patient age, tumor size, and nodal status. There was no significant relationship between lymphocytic immunity against tumor antigen and ER or PgR content of tumors, suggesting that patient lymphocyte immunity against tumor is independent of hormone receptor status. This is further evidence that lymphocyte immunity against tumor antigen status is an independent prognostic indicator that may be useful in the selection of a subset of node negative patients for adjuvant chemotherapy.     

Survival of premenopausal breast carcinoma patients in relation to menstrual cycle timing of surgery and estrogen receptor/progesterone receptor status of the primary tumor

BACKGROUND: Premenopausal breast carcinoma patients who undergo tumor excision during the follicular phase of their menstrual cycle may have a significantly worse prognosis than those whose tumors are excised in other phases of the menstrual cycle. METHODS: Outcome was determined in a series of 112 premenopausal women with operable breast carcinoma in relation to the timing of surgery within the menstrual cycle and the estrogen receptor (ER) and progesterone receptor (PR) status of their primary tumors as determined by immunohistochemistry. RESULTS: Those patients with ER positive tumors who underwent surgery in the early and luteal phase of the cycle had a significantly better survival than women with ER negative tumors (chi-square test = 15.56; P < 0.001). This also was true for PR status (chi-square test = 18.21; P < 0.001). After follicular phase surgery, tumor receptor status had no effect on overall survival. Patients with the best prognosis had ER/PR positive tumors excised on Days 0-2 and 13-32 but even those women with ER or PR negative tumors removed during the luteal phase of their menstrual cycle fared better than patients whose tumors were removed during the follicular phase. CONCLUSIONS: There was a better survival rate for patients with both ER/PR positive and negative tumors treated during the luteal phase of the menstrual cycle. This could be the result of progesterone acting on the surrounding peritumoral normal tissue, thereby exerting a straitjacket effect and improving cohesion of the primary carcinoma. Unopposed estrogen in the follicular phase of the cycle may enable more tumor emboli to escape and successfully establish micrometastases.     

CASP8 polymorphisms, estrogen and progesterone receptor status, and breast cancer risk

OBJECTIVES: This study was conducted to evaluate the potential role of CASP8 genetic polymorphisms in the etiology of breast cancer in a case-control study, Korea. METHODS: Incident breast cancer cases confirmed histologically (n = 1,599) were recruited from two hospitals in Seoul during 2001-2005. Control subjects (n = 1,536) were selected from the Health Examinee Cohort from Seoul and Gyeonggi Province surrounding Seoul, Korea. Three SNPs (D302H D > H, 5'-UTR C > T, and K337K G > A) were genotyped by the primer extension assay. The CASP8 D302H, which was not polymorphic in 48 samples, was excluded in further genotyping. Odds ratios and 95% confidential intervals (95% CIs) were estimated by unconditional logistic regression model adjusted for age at enrollment, education, age at first full-term pregnancy, cigarette smoking, and family history of breast cancer. RESULTS: The 5'-UTR T allele containing genotypes (CT/TT) were associated with an increased risk of breast cancer, compared with those with the CC genotype (OR = 1.13, 95% CI = 0.95-1.34; and OR = 1.48, 95% CI = 1.04-2.10, respectively; P-trend = 0.02). When stratified by the estrogen and progesterone receptor status, the association between the 5'-UTR T allele and breast cancer risk was prominent in ER(+) and PR(+) cases among pre-menopausal women (OR = 1.31, 95% CI = 1.00-1.72 and OR = 1.40, 95% CI = 1.06-1.85, respectively), whereas the association was found prominent in ER(-) or PR(-) cases (OR = 1.32, 95% CI = 0.93-1.87 and OR = 1.42, 95% CI = 1.04-1.94, respectively) among post-menopausal women. CONCLUSION: Our results thus suggest that the CASP8 5'-UTR C > T are associated with breast cancer risks and the effect may be modified by estrogen and progesterone receptor status.     

Effect of hormone replacement therapy on breast cancer risk: estrogen versus estrogen plus progestin

BACKGROUND: Hormone replacement therapy (HRT) given as unopposed estrogen replacement therapy (ERT) gained widespread popularity in the United States in the 1960s and 1970s. Recent prescribing practices have favored combination HRT (CHRT), i.e., adding a progestin to estrogen for the entire monthly cycle (continuous combined replacement therapy [CCRT]) or a part of the cycle (sequential estrogen plus progestin therapy [SEPRT]). Few data exist on the association between CHRT and breast cancer risk. We determined the effects of CHRT on a woman's risk of developing breast cancer in a population-based, case-control study. METHODS: Case subjects included those with incident breast cancers diagnosed over 4(1/2) years in Los Angeles County, CA, in the late 1980s and 1990s. Control subjects were neighborhood residents who were individually matched to case subjects on age and race. Case subjects and control subjects were interviewed in person to collect information on known breast cancer risk factors as well as on HRT use. Information on 1897 postmenopausal case subjects and on 1637 postmenopausal control subjects aged 55-72 years who had not undergone a simple hysterectomy was analyzed. Breast cancer risks associated with the various types of HRT were estimated as odds ratios (ORs) after adjusting simultaneously for the different forms of HRT and for known risk factors of breast cancer. All P values are two-sided. RESULTS: HRT was associated with a 10% higher breast cancer risk for each 5 years of use (OR(5) = 1.10; 95% confidence interval [CI] = 1.02-1.18). Risk was substantially higher for CHRT use (OR(5) = 1.24; 95% CI = 1.07-1.45) than for ERT use (OR(5) = 1. 06; 95% CI = 0.97-1.15). Risk estimates were higher for SEPRT (OR(5) = 1.38; 95% CI = 1.13-1.68) than for CCRT (OR(5) = 1.09; 95% CI = 0. 88-1.35), but this difference was not statistically significant. CONCLUSIONS: This study provides strong evidence that the addition of a progestin to HRT enhances markedly the risk of breast cancer relative to estrogen use alone. These findings have important implications for the risk-benefit equation for HRT in women using CHRT.